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ÄKTA start Operating Instructions

1. ar sassunEnEE ezEEE m EHH ae EEEE y i L PLL G SES00 Se ee s Ea ee a ee Ske00 CSE er ANNE amp PELE REECE HE BEONE ESC NEN ENNEN rE ETT TT rrr yg Tr A eee gt FHA 5 EESSERREEE ANEERDEE E BEE EEE eA oe PEE SEGG0 CRO SaSG08 PEs ee ees ee EEANN SOP 40 See EEREN oe O E A eee DRENE ie ee NDENE 95 HAHH HHHH HH AAE HHHH SEG8 SS ee nnn gees TE a EHS S008 See S008 SEDEN ee 7 Gia eee MN a 30 Temperature C 104 AKTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 3 Calibrations 5 3 6 Conductivity Monitor calibration Description Temperature C Conductivity mS cm AKTA start Operating Instructions 29 0270 57 AG 105 5 Prepare the system for a run 5 4 System performance 5 4 System performance Introduction This section describes the System performance method and how to perform and eval uate the System performance method system performance In this section This section contains the following subsections Section See page 5 4 1 System performance method 107 5 4 2 System performance method from AKTA start 109 5 4 3 System performance method from UNICORN start 114 5 4 4 Switch valve timing 117 106 AKTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 4 System performance 5 4 1 System performance met
2. 214 AKTA start Operating Instructions 29 0270 57 AG 7 Operations from UNICORN start 7 3 Evaluation 7 3 Evaluation Introduction The Evaluation module is used to manage and evaluate the results from chromatography runs Illustration of the Evaluation module The user interface of Evaluation is illustrated below g Evaluation GF 27 oo e e fal File Edit View Integrate Operations Tools Help Es Aa AL Peak Integrate We Edit Peak Table _ Default Results Recent Runs Find Resuts Open 5 a i UV 3 Conductivity 2 Gradient Concentration 4 System Flow O Pressure M Fraction Folder name System 4 Injection 4 Run Log H TEST PC mS cm 5 DefauttHome 18 5 00000000000001722538 a x AC GS Trap 0000000000 16 EJ AC HisTrap1 0000000000 15 AJ AC HisTrap2 0000000000 14 J AC Mab Protein A ID_nnnn nnni 13 fx AC Mab Protein G 1D_nnnn nnni 12 WLI DS HiTrap Frac Pump 002 1D_nnnn nnni 11 m DS HiTrap NoFrac Pump 0 1D_nnnn nnni 10 IGF 1 9 EGF 2 gs xj GF 3 7 ji A GF HiPrep Sephacryl 16_6 0000000000 5 AU IEX HiTrap DEAE 1 ID_nnnn nnni A Z HJ IEX HiTrap DEAE 2 ID_nnnn nnn j 3 g a IEX HiTrap DEAE 3 1D_nnnn nnni 4 5 O 3 5 1 1 0 Main features The main features of the Evaluation module are listed below e Open and view existing chromatogram results e Compare two curves or chromatograms e Perfor
3. KTA start Operating Instructions 29 0270 57 AG 13 1 Introduction 1 4 Associated documentation Access documentation from mobile units Scan the code using your mobile phone or tablet computer to access the product page for AKTA start Select documents to download under the Related Documents tab 14 AKTA start Operating Instructions 29 0270 57 AG 2 Safety instructions 2 Safety instructions About this chapter This chapter describes safety precautions and emergency shutdown procedures for the product The labels on the system and recycling procedures are also described In this chapter This chapter contains the following sections Section See page 2 1 Safety precautions 16 2 2 Labels 24 2 3 Emergency procedures 28 2 4 Recycling procedures 30 2 5 Declaration of Hazardous Substances DoHS 31 AKTA start Operating Instructions 29 0270 57 AG 15 2 Safety instructions 2 1 Safety precautions 2 1 Safety precautions Introduction AKTA start is powered by mains voltage and handles liquids that may be hazardous Before installing operating or maintaining the system you must be aware of the hazards described in this manual Follow the instructions provided to avoid personal injuries or damage to the equipment The safety precautions in this section are grouped into the following categories General precautions Using flammable liquids Personal protection Installing and moving the instrument Syste
4. End fractionation 1 Weigh fraction no 2 3 and 4 0 8 to 1 2g Max diff between fractions 0 19 End collection Weigh beaker 4 2 to 5 89 End gradient stay at 100 B Gradient Straight no nega tive dips 50 B Gradient level 45 to 55 B O B Re equilibration End 1 With Fraction collector 2 Without Fraction collector 3 UV 50 B UV 100 B Note Check all connec tions for leakage No leakages If the System performance method fails analyze the cause for the failure based on the acceptance criteria Perform the following actions e Re calibrate the failed module e Use buffer with proper composition e Clean the failed module or entire system Refer Chapter 8 Maintenance on page 217 for more details on cleaning e Carefully follow the test instructions e Repeat the System performance method until it passes AKTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 4 System performance 5 4 2 System performance method from AKTA start If wavy or fluctuations in gradient is observed then perform Switch valve timing optimization If the test fails after following the above actions replace the failed module AKTA start Operating Instructions 29 0270 57 AG 113 5 Prepare the system for a run 5 4 System performance 5 4 3 System performance method from UNICORN start 5 4 3 System performance method from UN
5. KTA start Operating Instructions 29 0270 57 AG Description Conductivity mea surement with the same buffer changes over time at constant tem perature Waves on the gra dient Possible cause Flow cell not clean Pump or Buffer valve malfuntion 9 Troubleshooting 9 2 Basic troubleshooting Action Clean the Conductivity flow cell see Section 8 5 Cleaning the Conductivity flow cell on page 229 Check that the Pump and the Buffer valve operate properly Refer to KTA start Maintenance Manual Switch valve tim ing not optimized Perform Switch valve timing to optimize the switch valve timing See Section 5 4 4 Switch valve timing on page 117 Temperature sen sor not calibrated Calibration solu tion 1 00 M NaCl not correctly pre pared Wrong absolute conductivity value Conductivity flow cell not calibrated Recalibrate the Conductivity flow cell See Section 5 3 Calibrations on page 88 Calibrate the temperature sensor See Section 5 3 Calibrations on page 88 Prepare a new calibration solution and recalibrate the Conductivity flow cell Ghost peaks in the gradient profile A charged sample was detected e g a protein Clean the Conductivity flow cell see Section 8 5 Cleaning the Conductivity flow cell on page 229 Air bubbles pass through the flow cell Check for loose tubing connections Non linear gradi ents or slow re s
6. 1 Place a precision thermometer in the path of the UV flow cell directly after the Conductivity flow cell and pump DM water through system with a flow rate of 0 5 ml min In the Settings and service screen 3 tap Conductivity to access the Con ductivity Monitor options Result The Conductivity screen opens Conductivity Calibration Configuration Advanced calibration In the Conductivity screen tap Advanced calibration Result The Advanced calibration screen opens Advanced calibration Setactualtemp ES 25 0 A ike Calibrate GEHC service only Cell resistance Ohms Calibrate sine gen cet a Note the temperature and type it into the Set Actual Temp field then tap Calibrate to carry out the temperature calibration Note Make sure that the temperature of the Conductivity flow cell has stabilized and measure the temperature of the calibration solution with a precision thermometer Note Calibrate Sine Gen is performed by a GE Service Engineer only when the Main board is replaced KTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 3 Calibrations 5 3 6 Conductivity Monitor calibration Calibrate the Conductivity flow cell Prequisites Calibration solution 1 00 M NaCl or 100 mS cm conductivity standard solution Follow the instructions below to calibrate the Conductivity flow cell Action In the Conductivity screen tap
7. The PEEK tubing should extend slightly 2 to 3 mm out of the tubing holder Make sure that the extended length of the tubing is short enough to avoid any collision with the test tubes during fractionation AKTA start Operating Instructions 29 0270 57 AG 147 5 Prepare the system for a run 5 8 Prepare the Fraction collector Step Action 4 Fit the tubing holder into the corresponding port on the Dispenser arm i e the outer or inner port is chosen according to the type of collection tubes inserted in the Bowl assembly 5 Gently move the arm to the dispensing position 148 AKTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 8 Prepare the Fraction collector Set the delay volume The delay volume represents the volume between the UV and the Fraction collector or the outlet that is used The delay volume settings are done to make sure that the fractions collected during fractionation correspond to the fractions indicated in the chromatogram When the Fraction collector is enabled the delay volume is collected in the first tube T1 and the elution volume is collected in the subsequent tubes Without the Fraction collector enabled the delay volume is collected in the collection beaker the total collected volume in the collection beaker will be delay volume elution volume As the delay volume is affected by the length and diameter of the tubing it should be set according to the tubing u
8. 3 From the KTA start home screen tap Method Run KTA start Method run Manual run Create method Settings and service In the Method run screen tap Prepare system Method run 2 Quick start Templates User defined Prepare system gp Q Select System performance method To initiate the method tap Select Select system method 2 System cleaning System performance method 2 2 AKTA start Operating Instructions 29 0270 57 AG 109 5 Prepare the system for a run 5 4 System performance 5 4 2 System performance method from AKTA start Step Action 4 Tick the check box in order to save the results on a USB memory stick and then tap Run to start the System Performance test Note Provide a unique file name Run parameters 2 System performance method yet ze PerfOl to USB S Q Den 1 1 Result The Run view screen will be displayed Run view UV absorbance 1 0 mAU Conductivity 25 0 mS cm Flow rate 3 0 ml min Graph Pressure 0 30 MPa Tube no 6 Edit run End Note e tis recommended not to edit any run parameters during a run to avoid the failure of the test e f required the System performance method run can be ended before it is completed tap End to abort the test 110 AKTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 4 System performance 5 4 2 System performance method from AKTA star
9. The Pump wash A method is used before the start of a new run or in case of changing the buffers During Pump wash A the flow is directed through Wash valve to Waste Note Pump wash A is performed at 10 ml min for 1 min through Buffer A port Pump wash A is important for preventing carryover and cross contamina tion among Buffers Samples It is recommended to first wash with DM water and then wash with the buffer of choice The Pump wash A method cannot be edited Requirements Required solutions are Instruction 126 e DM water e Buffer solution Follow the instructions below to perform a Pump wash Arun The Pump wash A proce dure is initiated from the Instrument Display Step Action i Immerse the buffer A inlet tubing in DM water or buffer 2 In the KTA start home screen tap Method run KTA start EA Create method Settings and service KTA start Operating Instructions 29 0270 57 AG Step 5 Prepare the system for a run 5 6 Run Prepare system methods 5 6 1 Pump wash A Action In the Method run screen tap Prepare system Method run 2 Quick start Templates User defined Prepare system Select Pump wash A and then tap Select to initiate the method Select system method 2 Pump wash A Pump wash B gt gt X Washout fractionation tubing X Column preparation 1 2 Result The following screen opens
10. 4 Installation 4 2 Transport KTA start and Frac30 AKTA start Operating Instructions 29 0270 57 AG 63 4 Installation 4 3 Unpack KTA start and Frac30 4 3 Unpack KTA start and Frac30 Introduction This section describes how to unpack AKTA start and Frac30 Note Save all the original packing material If the system has to be repacked for transportation or otherwise it is important that the system can be safely packed using the original packing material Unpack AKTA start Follow the instructions below to unpack the instrument CAUTION KTA start is filled with denaturated alcohol 50 C2H5OH ethanol at delivery The denatured alcohol mixture can be hazardous to humans if consumed Flush out the denaturated alcohol before assembling testing or integrating AKTA start into the intended process context Note AKTA start with packaging weighs about 12 kg No lifting equipment required one person can lift and move the instrument Step Action 1 Open the delivery box by cutting the adhesive tape at the top of the box 64 KTA start Operating Instructions 29 0270 57 AG 4 Installation 4 3 Unpack KTA start and Frac30 Step Action 2 Take out the document placed at the top of the package and read the Un packing Instructions Note Save the documents for future reference 3 Take out the box placed at the top of the package The box contains the accessories delivered with the i
11. KTA start home screen tap Method run KTA start 2 Create method Settings and service AKTA start Operating Instructions 29 0270 57 AG Step 5 Prepare the system for a run 5 6 Run Prepare system methods 5 6 2 Pump wash B Action In the Method run screen tap Prepare system Method run 2 Quick start Templates User defined Prepare system Q Select Pump wash B and then tap Select to initiate the method Select system method Pump wash A Pump wash B m Ra Washout fractionation tubing y Column preparation 1 2 Result The following screen opens Back Pump wash in progress UV absorbance 1 0 mAU Conductivity 25 0 mS cm Flow rate 3 0 ml min Graph Pressure 0 30 MPa Tube no 6 Hold E If required the Pump wash B run can be ended before it is completed by tapping End to stop the wash in advance When the run is completed tap Exit to close the Pump wash B screen Run ended a UV absorbance 1 0 mAU Conductivity 25 0 mS cm Flow rate 0 0 ml min Graph Pressure 0 30 MPa Tube no 6 Exit KTA start Operating Instructions 29 0270 57 AG 129 5 Prepare the system for a run 5 6 Run Prepare system methods 5 6 3 Washout fractionation tubing 5 6 3 Washout fractionation tubing Introduction The Washout fractionation tubing method is used to clean the fractionation tubing It is recommended while collecting the fractions without Fract
12. Pump wash in progress UV absorbance 1 0 mAU Conductivity 25 0 mS cm Flow rate 3 0 ml min Graph Pressure 0 30 MPa Tube no 6 Hold RRR ena Note If required the Pump wash A run can be ended before it is completed by tapping End to stop the run When the run is completed tap Exit to close the Pump wash A screen Run ended g UV absorbance 1 0 mAU Conductivity 25 0 mS cm Flow rate 0 0 ml min Graph Pressure 0 30 MPa Tube no 6 Exit AKTA start Operating Instructions 29 0270 57 AG 127 5 Prepare the system for a run 5 6 Run Prepare system methods 5 6 2 Pump wash B 5 6 2 Pump wash B Introduction The Pump wash B method is used before the start of a new run or in case of changing the buffers During Pump wash B the flow is directed through Wash valve to Waste Note e Pump wash B is performed at 10 ml min for 1 min through Buffer B port e Pump wash Bis important for preventing carryover and cross contamina tion among buffers and samples e tis recommended to first wash with DM water and then wash with the buffer of choice e The Pump wash B method cannot be edited Requirements Instruction 128 Required solutions are e DM water e Buffer solution Follow the instructions below to perform a Pump wash B run The Pump wash B proce dure is initiated from the Instrument Display Step Action i Immerse the buffer B inlet tubing in DM water or buffer 2 In the
13. When replacing system tubing use the original inner diameter and length to ensure that the correct delay volumes are maintained Inlet and outlet tubing may be shortened if required 5 Put the labels back on the new tubing 6 Mount the connectors on the tubing For fingertight connectors e Slide the connector onto the tubing For tubing connectors 1 8 e Slide the connector onto the tubing e Slide the ferrule onto the tubing with the thick end towards the end of the tubing 7 Insert the tubing with connector into the port Make sure to insert the tubing all the way into the bottom of the port 8 Tighten the connector fully AKTA start Operating Instructions 29 0270 57 AG 241 9 Troubleshooting 9 Troubleshooting About this chapter This chapter describes troubleshooting and corrective actions for AKTA start In this chapter This chapter contains the following sections Section See page 9 1 Introduction to troubleshooting 243 9 2 Basic troubleshooting 245 9 3 System error report 256 242 AKTA start Operating Instructions 29 0270 57 AG 9 Troubleshooting 9 1 Introduction to troubleshooting 9 1 Introduction to troubleshooting Introduction The sections in this chapter describe the basic troubleshooting for AKTA start and include a general checklist that should be completed prior to the troubleshooting work How to generate a System error report for service purposes is also presented For replacement
14. e Toggle as needed to set which valve positions are open Sample valve set as Buffer or Sample so that the flow is delivered from either the buffer inlets or the sample inlet Washvalve set as Column or Waste to direct the flow to either the column or to waste Outlet valve set as Collection or Waste to direct the flow to either the Fraction collector or to waste AKTA start Operating Instructions 29 0270 57 AG 167 6 Operations from the Instrument Display 6 3 Perform a manual run 6 3 2 Monitor and control the run Step Action e Tick the Autozero UV checkbox if a UV baseline to zero is required e Tick the Set event mark checkbox if you need to set an event mark in the chromatogram e After you have set the run parameters tap Execute to implement the changes Pause the run Follow the instruction below to pause an ongoing run Step Action 1 In the Run view screen tap Pause to temporarily pause the run by stopping the Pump Run view UV absorbance 1 0 mAU Conductivity 25 0 mS cm i Flow rate 3 0 ml min Graph Pressure 0 30 MPa Tube no 6 emr lt 2 To continue the run tap Continue Run paused UV absorbance 1 0 mAU Conductivity 25 0 mS cm Flow rate 3 0 ml min Graph Pressure 0 30 MPa Tube no 6 Edit run End Note The run is paused and there is no flow of liquid in the flow path as the Pump is stopped 168 KTA start Opera
15. s50 yom mi ta iC Resul to USB esultOl 1 1 Run KTA start Operating Instructions 29 0270 57 AG 187 6 Operations from the Instrument Display 6 4 Perform a method run 6 4 4 User defined methods 188 Step Action Set the run parameters e Select the mode of sample injection in the Sample from field to be ap plied via Pump or via Loop For detailed instructions on sample applica tion refer to Section 5 7 Sample application on page 135 e Sample volume the volume of sample to be loaded in to the column e Tick the checkbox Save Result to USB if you want to save the result The result file name can be edited by setting the digits in the range 00 to 99 e Tap Run to start the selected method Result The Run view screen will be displayed 4 Run view UV absorbance 1 0 mAU Conductivity 25 0 mS cm s Flow rate 3 0 ml min Graph Pressure 0 30 MPa Tube no 6 e a Edit run Pause End In the Run view screen the following options are available to monitor and control the ongoing run for details see Section 6 3 2 Monitor and control the run on page 165 Graph to view the chromatogram Edit run to change any run parameters in the current run Hold to hold the current run Pause to pause the current run End to end the run before it is completed Note The USB memory stick stores the result files which can be viewed using UNICORN start Also a bmp file is generated wh
16. 1 Introduction 1 2 Important user information 1 2 Important user information Read this before operating the product All users must read the entire Operating Instructions before installing operating or maintaining the product Always keep the Operating Instructions at hand when operating the product Do not operate the product in any other way than described in the user documentation If you do you may be exposed to hazards that can lead to personal injury and you may cause damage to the equipment Intended use KTA start is a liquid chromatography system used for preparative purification of proteins at laboratory scale The system can be used for a variety of research purposes to fulfill the needs of the users in the academia and in the life sciences industry AKTA start is intended for research use only and shall not be used in any clinical proce dures or for diagnostic purposes Prerequisites In order to follow this manual and use the system in the manner it is intended it is im portant that e you understand the concepts of liquid chromatography e you have read and understood the Safety instructions chapter in the KTA start Operating Instructions AKTA start Operating Instructions 29 0270 57 AG 7 1 Introduction 1 2 Important user information Safety notices This user documentation contains safety notices WARNING CAUTION and NOTICE concerning the safe use of the product See definitions b
17. Elution volume volume of buffer needed to elute the protein from the column Fractionation volume volume of fractions to be collected when the Fraction collector is enabled e Tap Run to start the run Result The Run view screen will be displayed Note Conc B is not applicable for GF DS as elution takes place only with a single buffer buffer A Note The run begins with a default pump wash Pump wash is performed with buffer A at 10 ml min for 30 sec Edit run is disabled when the pump wash is in progress During pump wash the flow is directed through Wash valve to Waste 186 AKTA start Operating Instructions 29 0270 57 AG 6 Operations from the Instrument Display 6 4 Perform a method run 6 4 4 User defined methods 6 4 4 User defined methods Start a run Follow the instruction below to start a run based on a user created or USB imported method Note Make sure that a USB memory stick is connected to the instrument If the instrument does not detect a USB memory stick the result will not be saved Step Action 1 Method run 2 Quick start Templates User defined Prepare system In the Method run screen tap User defined to access the user created methods Select method AC01 DS06 Lak EZ e Tapa radio button to select a user method to run e To continue with the selected user method tap Select Run parameters Sample from Pump Loop Sample volume v
18. Introduction This section describes how to start a manual run by configuring the run parameters from the instrument display and how to control an ongoing run In this section This section contains the following subsections Section See page 6 3 1 Manual run 163 6 3 2 Monitor and control the run 165 162 AKTA start Operating Instructions 29 0270 57 AG 6 3 1 Start a run 6 Operations from the Instrument Display 6 3 Perform a manual run 6 3 1 Manual run Manual run Follow the instruction below to start a manual run Note Make sure that a USB memory stick is connected to the instrument If the instrument does not detect a USB memory stick the result will not be saved Step Action 1 In the AKTA start home screen tap Manual run to access the run parameters for a manual run AKTA start 2 Settings and service Tap the forward arrows to access additional run parameters 2 Manual run Flow rate Zam ml min Pressure limit v ym MPa Conc B ym 6 A Save Result ManRes0O1 1 2 to USB lt lt Vv e Setrun parameters Flow rate flow rate ml min Pressure limit pressure limit MPa Conc B buffer B concentration Use up down arrows to set the values or use the numpad to type in the values e Tickthe checkbox Save Result to USB if you want to save the result The result file name can be edited by setting the digits in the range 00 to 99 e Tap Run if no other parameters need
19. Note If required the Column preparation run can be ended before it is completed by tapping End to stop the run i When the run is completed tap Exit to close the Column preparation screen 134 KTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 7 Sample application 5 7 Sample application Sample application The table below shows the different modes of sample application available for AKTA start The sample application technique can be selected from the Instrument display in the Run parameters screen or from UNICORN start For details see Chapter 6 Operations from the Instrument Display on page 155 and UNICORN start 1 0 User Manual Sample volume Sample application Loop type 25 ulto 5ml via Loop Sample loop 10 ml to 150 ml via Loop Superloop 10 ml Superloop 50 ml Superloop 150 ml via Pump from the Sample valve port I Sample Note Make sure to load only the recommended volume of sample in the column to obtain good results For details see the Column instructions KTA start Operating Instructions 29 0270 57 AG 135 5 Prepare the system for a run 5 7 Sample application Injection valve description The Injection valve enables the application of a sample onto the column from a sample loop connected to the valve The illustration below shows the different positions of the Injection valve The Injection valve position can be changed manually by turning the le
20. Note Step Make sure that the UV flow cell is assembled tightly and the inlet and outlet tubes are fitted tightly to the UV Monitor No stray light should enter the UV flow cell Action 98 Immerse the buffer inlet tubing in DM water Flush the UV flow cell with DM water using the Pump Make sure that there are no air bubbles in the UV flow cell Note Flush thoroughly with DM water to make sure that the flow cell is clean In the AKTA start home screen tap Settings and service to access the in strument modules then tap Next Next Result The Settings and service Screen 3 opens Settings and service EJ Display Conductivity KTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 3 Calibrations 5 3 5 UV Monitor calibration Step Action 4 In the Settings and service screen tap UV to access the UV Monitor options Result The UV screen opens UV 2 UV LED calibration Flow cell path length Diagnostics Configuration Back 5 In the UV screen tap UV LED calibration For a description of the UV calibra tion parameters refer to the KTA start Maintenance Manual Result The UV LED Calibration screen opens UV LED calibration 2 Light Strength T amb oo ec Calibrate Save Back om 6 e Setthe Light strength value to 500 using up down arrows or type in the value e Tap Calibrate A confirmation screen opens Select OK to
21. On Site Service Health amp Safety Declaration Form Service Ticket To make the mutual protection and safety of GE service personnel and our customers all equipment and work areas must be clean and free of any hazardous contaminants before a Service Engineer starts a repair To avoid delays in the servicing of your equipment please complete this checklist and present it to the Service Engineer upon arrival Equipment and or work areas not sufficiently cleaned accessible and safe for an engineer may lead to delays in servicing the equipment and could be subject to additional charges Please review the actions below and answer Yes or No Provide explanation for any No answers in box below Instrument has been cleaned of hazardous substances Please rinse tubing or piping wipe down scanner surfaces or otherwise ensure removal of any dangerous residue Ensure the area around the instrument is clean If radioactivity has been used please perform a wipe test or other suitable survey Adequate space and clearance is provided to allow safe access for instrument service repair or installation In some cases this may require customer to move equipment from normal operating location prior to GE arrival Consumables such as columns or gels have been removed or isolated from the instrument and from any area that may impede access to the instrument All buffer waste vessels are labeled Excess containers h
22. The Settings and service Screen 1 opens Settings and service Fraction collector Pressure sensor Pump System 1 3 Next KTA start Operating Instructions 29 0270 57 AG 79 4 Installation 4 5 Install KTA start Step Action e Inthe Settings and service screen tap Fraction collector Result The Fraction collector screen opens Fraction collector Back A e Inthe Fraction collector screen tap Enable Frac to enable the connec tion of the Fraction collector Result The following screen will be displayed Fraction collector Disable Frac unlog Back Note The power on status of the Fraction collector is indicated by the white LED on the front of Frac30 Connect a computer to AKTA start Note Before connecting the computer to AKTA start install the UNICORN start soft ware on the computer Refer to the UNICORN start 1 0 User Manual Follow the instructions below to connect a UNICORN start computer to AKTA start 80 AKTA start Operating Instructions 29 0270 57 AG 4 Installation 4 5 Install AKTA start Step Action it Connect power to the computer and monitor and then switch on the com puter and AKTA start Result The instrument displays the AKTA start home screen AKTA start 2 Create method Settings and service 2 Connect the PC Connection Cable between the connector marked as PC Connection on the back
23. This chapter provides the necessary instructions to enable users to unpack and install KTA start and Frac30 Read the entire Installation chapter before starting to install KTA start In this chapter This chapter contains the following sections Section See page 4 1 Space requirements 60 4 2 Transport AKTA start and Frac30 62 4 3 Unpack AKTA start and Frac30 64 4 4 Accessories package 74 4 5 Install AKTA start 15 AKTA start Operating Instructions 29 0270 57 AG 59 4 Installation 4 1 Space requirements 4 1 Space requirements Benchtop setup The illustration below shows the space requirements recommended for AKTA start 60 AKTA start Operating Instructions 29 0270 57 AG 4 Installation 4 1 Space requirements Equipment dimensions AKTA start Frac30 e85 mm 280 mm AKTA start Operating Instructions 29 0270 57 AG 61 4 Installation 4 2 Transport AKTA start and Frac30 4 2 Transport KTA start and Frac30 Equipment weight AKTA start with packaging Frac30 with packaging Handling the delivery boxes AKTA start and Frac30 are packed in two separate boxes GE Healthcare Life Sciences Frac30 To transport the delivery boxes containing the instrument and Fraction collector use a hand truck suitable for light weight packages However each box can be lifted by 1 person without the help of any lifting equipment 62 AKTA start Operating Instructions 29 0270 57 AG
24. menu for KTA start only accessible from the Instrument Display KTA start Maintenance Manual Detailed instrument and module descrip tions including instructions needed to maintain and troubleshoot KTA start KTA start System Cue Card A condensed guide to prepare and run chromatographic techniques on KTA start KTA start Maintenance Cue Card 12 A condensed guide to handling routine maintenance operations and trou bleshooting KTA start KTA start Operating Instructions 29 0270 57 AG 1 Introduction 1 4 Associated documentation From the Help menu in UNICORN start or on the UNICORN start DVD the following user documentation is available Documentation Main contents UNICORN start 1 0 User Manual Overview and detailed descriptions of the system control software designed for KTA start which includes process pic ture map for real time monitoring method editor evaluation and adminis tration modules UNICORN start Online Help Dialog descriptions for UNICORN start only accessible from the Help menu Data files application notes and user documentation on the web To order or download data files application notes or user documentation see the instruc tion below Step Action 1 Go to www gelifesciences com AKTA 2 Click KTA Lab Scale Systems 3 Select KTA start from the list 4 Click Related Documents 5 Select to download the chosen literature
25. the start of the run Fraction collector is not connected to the instrument Connect the Fraction collector to KTA start Fraction collector option is not en abled Enable the Fraction collector option from the Settings and service screen 251 9 Troubleshooting 9 2 Basic troubleshooting Buffer valve Sample valve Wash valve Outlet valve Description External leakage Possible cause Tubing connec tions Action Check the tubing connections Tighten or replace if required Internal leakage Internal parts may be worn Replace valve Refer to KTA start Maintenance Manual High back pres sure Flow path not clean Clean the system flow path See Sec tion 8 3 Cleaning the system flow path on page 222 Valve not switch ing position Injection valve 252 Description The valve switch ing to wrong posi tion Internal parts may be worn Possible cause Valve parts incor rectly assembled after replacement Replace valve Refer to KTA start Maintenance Manual If the valve does not function even after replacement contact a GE Service Engi neer Action Make sure that the marking etched on the valve aligns correctly with the same marking on the instrument Refer to KTA start Maintenance Manual External leakage Tubing connec tions Check the tubing connections Tighten or replace if required Internal leakage Inte
26. 1 16 female 1 16 female for 18 3855 01 1 16 o d tubing titanium Union Valco F F 11 0003 39 Fill port 18 1127 66 Tubing Inlet tubing Kit KTA start 29 0240 32 Complete tubing kit KTA start 29 0240 34 PEEK tubing i d 0 75 mm 1 16 18 1112 53 PEEK tubing i d 1 0 mm 1 16 18 1115 83 PEEK tubing 2 m i d 0 5 mm o d 1 16 18 1113 68 Cables Mains cable 115 V 19 2447 01 Mains cable 220 V 19 2448 01 Cable Assy OTH USB 29 0240 36 212 KTA start Operating Instructions 29 0270 57 AG Miscellaneous 10 Reference information 10 5 Ordering information Accessory description Inlet filter assembly Code no 18 1113 15 Inlet filter set 10 Filters Nets 18 1114 42 Screw lid GL45 kit KTA 11 0004 10 Tubing cutter 18 1112 46 Column clamp o d 10 to 21 mm 20 9503 19 Short column holder 18 1113 17 T Slot holders 29 0240 38 Buffer tray KTA start 29 0240 39 Accessory Box 29 0240 37 Operating Instructions printed 29 0270 57 Maintenance Manual printed 29 0603 08 Injection kit 18 1110 89 Software UNICORN start DVD license access code and manual package 29 0187 51 Frac30 KTA start spare parts Frac30 Assembly 29 0230 51 Drive sleeve 19 6067 02 Tubing holder Bowl Assembly Frac30 18 6464 01 29 0240 45 Cable Assembly Frac30 Packaging Kit for AKTA start 29 0240 65 Co
27. 200 664 3 CIP RPC Cell sepa ration DTT 100 mM 3483 12 3 222 468 7 Reducing agent DTE 100 mM 6892 68 8 229 998 8 Reducing agent TCEP Tris 2 Car 100 mM 51805 45 9 Reducing agent boxyethy Phosphine EDTA 100 mM 6381 92 6 205 358 3 Buffer additive Ethanol 96 64 17 5 200 578 6 Storage Long term use Ethylene glycol 30 112 60 7 203 989 9 Buffer additive Glycerol 30 56 81 5 200 289 5 Buffer additive Glycine 0 5 M 56 40 6 200 272 2 Cleaning of MAb binding media KTA start Operating Instructions 29 0270 57 AG 10 Reference information 10 2 Chemical resistance Chemical Guanidine hydrochloride Hydrochloric acid Concentration 6M CAS no EEC no 50 01 1 200 002 3 7647 01 0 231 595 7 Usage Denaturing of proteins CIP Imidazole 288 32 4 206 019 2 Affinity Isopropanol 67 63 0 200 661 7 CIP Methanol 67 56 1 200 659 6 RPC CIP Mercaptoethanol 60 24 2 200 464 6 Reducing agent Potassium phosphate basic 16788 57 1 231 834 5 Buffer SDS 151 21 3 205 788 1 Detergent Sodium chloride 7647 14 5 231 598 3 CIP Sodium hydroxide NaOH 1310 73 2 215 185 5 CIP Sodium sulphate 7757 82 6 231 820 9 Buffer Triton x 100 Tween 20 9002 93 1 9005 64 5 500 018 3 Detergent Detergent Urea Wet side and painting 57 13 6 200 315 5 Buffer additive denaturing
28. 5 10 Starting a run 153 KTA start Operating Instructions 29 0270 57 AG 83 5 Prepare the system for a run 5 1 Flow path overview Tel Flow path overview Illustration of the flow path 84 The illustration below shows the flow path for KTA start The flow path contains Pump Mixer Valves and UV Conductivity and Pressure monitors The individual instrument modules are presented in the table below For a detailed description of the modules see the KTA start Maintenance Manual Description Description Buffer valve Injection valve manual Mixer Column Sample valve UV Monitor Pump Conductivity Monitor Pressure sensor Outlet valve Wash valve Fraction collector AKTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 1 Flow path overview Inlet and outlet tubing AKTA start is delivered with the entire flow path assembled and prefilled with storage solution 50 Ethanol Details on the tubing types used along the flow path are presented in Chapter 10 Reference information on page 258 The table below lists the tubing connected to the instrument Prepare the system for a run by connecting inlet and outlet tubing to the valve ports marked with orange arrows Module Buffer valve Tube connection Port Buffer A Scope of use Inlet tubing for buffer A Port Il Outlet tubing to the Mixer Port III Buffer B Inlet tubing for buffer
29. AG 8 Maintenance 8 7 Storage of the instrument Step Action 4 Tap Edit run and set the run parameters as indicated below Result The sample inlet tubing and the outlet tubing for fraction collection will be flushed with DM water Parameter Setting Select Buffer Sample Sample Outlet valve Collection 5 Pump 20 ml of DM water through the system 6 End the run and leave the system filled with DM water during the storage period Long term storage If the system is not going to be used for more than 4 days follow the instructions below to prepare the system for long term storage Note Before cleaning the flow path remove the column and re connect the flow path See Section 8 3 1 Disconnect the column on page 223 Step Action 1 Immerse the buffer and the sample inlet tubing in 20 ethanol ra Start a manual run and pump 20 ml of 20 ethanol through the system Use the same run parameters recommended for the short term storage procedure 3 Edit the run and set the run parameters to clean the sample inlet tubing and the outlet tubing for fraction collection 4 Pump 20 ml of 20 ethanol through the system 5 End the run and leave the system filled with 20 ethanol during the storage period AKTA start Operating Instructions 29 0270 57 AG 237 8 Maintenance 8 8 Replacement of tubing and filters 8 8 Replacement of tubing and filters Introduction This section describes how to replace tubing and
30. Allows the user to perform calibra tion set the delay volume and diag nostics and or troubleshooting The menu covers the following modules Fraction collector Fraction col System FT Pressure sensor fat Next Pump Settings and service System System Settings and service Screen 2 Allows the user to perform diagnos tics and or troubleshooting for the modules Sample valve Wash valve 2 3 Settings and service Back Outlet valve Settings and service Screen 3 Allows the user to perform calibra tion diagnostics configuration and or troubleshooting for the mod ules pw Main board Main board UV Settings and service UV Display Display ry isplay Display Conductivity AKTA start Operating Instructions 29 0270 57 AG 51 3 System description 3 3 Instrument Display 3 3 4 Description of Settings and service Settings and service Screen 1 The options available for the modules displayed on Settings and service Screen 1 are briefly described below Module Function Fraction collector Fraction collector Enable Frac Disable Frac Acti vates Deactivates the Fraction col lector connection Diagnostics Performs a feed tube isable Frac test that checks if the Bowl assembly rotates correctly Performs a Home o Rg test to verify that the Bowl assembly returns to Home position Fraction collector e
31. Description of Method run Method run options The display option Method run allows the user to run methods based on Quick start techniques or predefined method templates user created methods and run predefined methods such as Pump Wash and System cleaning Detailed instructions to run methods are presented in Section 6 4 Perform a method run on page 170 When Method run is selected further options are displayed in the Method run screen Method run Quick start Templates User defined Prepare system The method types available under Method run are briefly described below Method Description Quick start 1 3 Displays the Quick start templates available with AKTA start Quick start ee For description of the Quick start ACstep 1 mi HiTrap methods see Section 6 4 2 Quick gt Seek rier E5 start on page 173 gt p g DSS mlHiTrap DS 53 mi HiPrep 1 3 Back tt EZE Quick start 2 3 Quick start EXstep 1 ml HiTrap C IEX step 5 ml HiTrap gt K C IEX gradient 1 ml HiTrap O IEX gradient 5 ml HiTrap 2 3 Back Q Select AKTA start Operating Instructions 29 0270 57 AG 47 3 System description 3 3 Instrument Display 3 3 2 Description of Method run Method Description Quick start 3 3 Quick start 2 D GF 16 60 HiPrep K 3 3 Back 2 EZS Templates Displ
32. Instrument Display all four touch buttons will be highlighted The run is interrupted immediately Data generated up to 10 seconds before the power failure can be recovered Note The UNICORN start client may close down during a temporary overload of the processor This may appear as a computer failure The run continues and you can restart the UNICORN start client to regain control KTA start Operating Instructions 29 0270 57 AG 29 2 Safety instructions 2 4 Recycling procedures 2 4 Recycling procedures Introduction This section describes the procedures for disposal and recycling of AKTA start Decommissioning of the equipment AKTA start must be decontaminated before decommissioning Follow local regulations for scrapping of the equipment Disposal general instructions When taking AKTA start out of service the different materials must be separated and recycled according to national and local environmental regulations Recycling of hazardous substances AKTA start contains hazardous substances Detailed information is available from your GE representative Disposal of electrical components Waste comprising electrical and electronic equipment must not be disposed of as un sorted municipal waste and must be collected separately Please contact an authorized representative of the manufacturer for information concerning the decommissioning of equipment 30 AKTA start Operating Instructions
33. Make sure to use recommended length and ID of the PEEK tubing from UV to Outlet valve to avoid incorrect calculation of the delay volume KTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 9 Cold room operations 5 9 Cold room operations Introduction When purifying biomolecules that are temperature sensitive chromatography runs are performed in a cold room Preparation Follow the instructions below to prepare the instrument for a run in the cold room Step Action 1 Place AKTA start in the cold room 2 If a UNICORN start computer is connected to the instrument leave the computer outside the cold room 3 Allow the instrument to stabilize at the temperature of the cold room 4 Tighten all connections and pump DM water through the system to check for leaks Tighten any leaking connector Starting a run Before starting a run make sure that the temperature of the buffers has reached the temperature set in the cold room Note The measured temperature of the system is the temperature in the Conduc tivity flow cell which can differ from the ambient temperature Removal from the cold room Follow the instructions below to remove the instrument from the cold room Step Action 1 Switch off the instrument and disconnect the power cable before moving the instrument out of the cold room 2 Loosen all connections to prevent them sticking when the system returns to roo
34. PEE E A EEO PAE O AE EEE EE 279 KTA start Operating Instructions 29 0270 57 AG 1 Introduction 1 Introduction About this chapter This chapter contains important user information description of safety notices regula tory information intended use of AKTA start and list of associated documentation In this chapter Section See page 1 1 About this manual 6 1 2 Important user information f4 1 3 Regulatory information 9 1 4 Associated documentation 12 KTA start Operating Instructions 29 0270 57 AG 5 1 Introduction 1 1 About this manual 1 1 About this manual Purpose of this document The Operating Instructions provide you with the instructions needed to install operate and maintain AKTA start in a safe way Nomenclature conventions The nomenclature used in this manual are explained in the table below Concept AKTA start Explanation The instrument Frac30 The Fraction collector UNICORN start The software installed on a computer KTA start System Typographical conventions The entire liquid chromatography system including instrument Fraction collector and software Software items are identified in the text by bold italic text A colon separates menu levels thus File Open refers to the Open command in the File menu Hardware items are identified in the text by bold text for example Buffer valve KTA start Operating Instructions 29 0270 57 AG
35. Prepare the system for a run 5 5 Connect a column Step Action 3 Fix the column to the column holder 4 Connect the 0 75 mm i d PEEK tubing from the Injection valve port 1 to the column head JECTION VALVE AKTA start Operating Instructions 29 0270 57 AG 123 5 Prepare the system for a run 5 5 Connect a column Step Action 5 Connect the 0 75 mm i d PEEK tubing from the bottom of the UV Monitor to the bottom of the column Note The 0 75 mm i d PEEK tubing should not be disconnected from the UV Monitor inlet when the column is removed See Section 8 3 1 Disconnect the column on page 223 INJECTION VALVE Note Do not overtighten when connecting columns Overtightening might break the connectors or squeeze the tubing and thereby obstructing the flow 124 AKTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 6 Run Prepare system methods 5 6 Run Prepare system methods Introduction This section describes how to prepare the flow path and the column before starting a chromatography run In this section This section contains the following subsections Section See page 5 6 1 Pump wash A 126 5 6 2 Pump wash B 128 5 6 3 Washout fractionation tubing 130 5 6 4 Column preparation 133 KTA start Operating Instructions 29 0270 57 AG 125 5 Prepare the system for a run 5 6 Run Prepare system methods 5 6 1 Pump wash A 5 6 1 Pump wash A Introduction
36. Pressure limit pressure limit MPa Run volume run volume ml Run parameters Column volume 10 ml Flow rate v 10 Ze ml min Pressure limit v 030 tom MPa Run volume v so am mi amp Run Tap Run to initiate the method KTA start Operating Instructions 29 0270 57 AG 131 5 Prepare the system for a run 5 6 Run Prepare system methods 5 6 3 Washout fractionation tubing Step Action Result The following screen opens Frac tube wash in progress UV absorbance 1 0 mAU Conductivity 25 0 mS cm Flow rate 3 0 ml min Graph Pressure 0 30 MPa Tube no 6 Edit run Hold Pause f End Note If required the Washout fractionation tubing run can be ended before it is completed by tapping End to stop the washout in advance 8 When the run is completed tap Exit to close the Washout fractionation tubing screen Run ended 2 UV absorbance 1 0 mAU Conductivity 25 0 mS cm Flow rate 0 0 ml min Graph Pressure 0 30 MPa Tube no 6 Exit 132 KTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 6 Run Prepare system methods 5 6 4 Column preparation 5 6 4 Column preparation Introduction The Column preparation method is used to prepare a new column or to equilibrate the column It is recommended to equilibrate columns before starting a new run Requirements Required solution is e Buffer solution Instructions Follow the
37. Safety instructions 2 2 Labels Safety symbols The following safety symbols are used in the labels Meaning Warning Do not use KTA start before reading the KTA start Operating Instructions Do not open any covers or re place parts unless specifically stated in the user docu mentation The system complies with the requirements for electro magnetic compliance EMC in Australia and New Zealand Ce The system complies with applicable European direc tives ly LISTED Intertek Labels concerning hazardous substances This symbol indicates that KTA start has been certified by a Nationally Recognized Testing Laboratory NRTL NRTL means an organization that is recognized by the US Occupational Safety and Health Administration OS HA as meeting the legal requirements of Title 29 of the Code of Federal Regulations 29 CFR Part 1910 7 The following symbols on the labels concern hazardous substances Meaning This symbol indicates that electrical and electronic equipment must not be disposed of as unsorted munic ipal waste and must be collected separately Please contact an authorized representative of the manufac turer for information concerning the decommissioning of equipment 26 KTA start Operating Instructions 29 0270 57 AG 2 Safety instructions 2 2 Labels Meaning This symbol indicates that the product contains haz ardous materials in excess of the limits
38. agent Long term use Chemical Concentration CAS no EEC no Decon 90 10 1310 58 3 215 181 3 Ethanol 20 64 17 5 200 578 6 Hydrochloric acid 0 1M 7647 01 0 231 595 7 lsopropanol 70 67 63 0 200 661 7 Triton x 100 1 9002 93 1 Tween 20 1 9005 64 5 500 018 3 266 AKTA start Operating Instructions 29 0270 57 AG 10 Reference information 10 2 Chemical resistance Chemical Concentration CAS no EEC no Spray with commercial house cleaning 5 N A detergent Display Chemical Concentration CAS no EEC no Aqueous buffers pH 2 to pH 12 N A Decon90 1310 58 3 215 181 3 Ethanol 64 17 5 200 578 6 Hydrochloric acid 7647 01 0 231 595 7 Isopropanol 67 63 0 200 661 7 Sodium chloride 7647 14 5 231 598 3 Sodium hydroxide i 1310 73 2 215 185 5 Triton X 100 9002 93 1 Tween 20 9005 64 5 500 018 3 Spray with commercial house cleaning N A detergent KTA start Operating Instructions 29 0270 57 AG 267 10 Reference information 10 3 Literature 10 3 Literature For further information related to KTA start refer to the following 268 AKTA start Maintenance Manual UNICORN start 1 0 User Manual AKTA start System Cue Card AKTA start Maintenance Cue Card Data file AKTA start Operating Instructions 29 0270 57 AG 10 Reference information 10 4 Health and Safety Declaration Form 10 4 Health and Safety Declaration Form On site service
39. b d j Mfg Yeor 2013 Max Power 95VA y Sym O S an Specl Fuse FSAL250V AX OCEA fication TE GE Healthcare Bio Sciences AB Made in Sweden 75184 Uppsala 36 d e CAN Dacca ikeaet aretati telah catia ita acc he adeat AD Sweden Conforms te ANSIUL Sad 61010 1 Ceit 1o CANICSA Sad 22 2 Ne 61010 1 Do not open any covers on the instrument This will void the warranty La rupture du sceau annule la garantie Warranty void if seal is broken Keep the pump cover open when not using the system Open the pump cover after you switch off the instrument 24 KTA start Operating Instructions 29 0270 57 AG 2 Safety instructions 2 2 Labels Description Placement Caution Consult the Operating Instructions before using the system Pinch hazard Switch off the Pump before tubing loading Label on Frac30 The illustration below shows the labels that are attached to Frac30 Description Placement 29023051 Frac30 Instrument label including Code no 29002975 Voltage 24 v 3 ia r safety symbols and speci Protection Closs 1p21 AELOC ED fication CRT GE Healthcore Bio Sciences AB Made in Sweden 1 Sweden 36 751 84 Uppsala Do not open any covers on the instrument This will void the warranty La rupture du sceau annule la garantie Warranty void if seal is broken AKTA start Operating Instructions 29 0270 57 AG 25 2
40. cleaning and then tap Select to initiate the method For de tailed instructions see Section 6 4 5 Prepare system methods on page 189 Select system method System cleaning C System performance method e Perform the operations presented on the display 1 Disconnect the column 2 Connect Injection valve to UV with suitable tubing and connectors 3 Place the buffer tubings A amp B in cleaning solution e Tap Continue to start System cleaning System cleaning Please follow instructions 1 Disconnect the column 2 Connect injection valve to UV with suitable tubing and connectors 3 Place the buffer tubings A amp B in cleaning solution When completed press Continue AKTA start Operating Instructions 29 0270 57 AG 225 8 Maintenance 8 3 Cleaning the system flow path 8 3 2 System cleaning 226 Step Action i When the run is completed tap Exit to close the System cleaning screen Note Do not leave NaOH in the instrument in order to avoid damage of the system Wash the complete flow path with water Check the pH after a wash with water to make sure a complete removal of NaOH System cleaning in progress UV absorbance 1 0 mAU Conductivity 25 0 mS cm Flow rate 3 0 ml min f Graph Pressure 0 30 MPa Tube no 6 Edit run End AKTA start Operating Instructions 29 0270 57 AG 8 Maintenance 8 4 Cleaning the UV flow cell 8 4 Cleaning the UV flow cell Maintenance interv
41. connectors and how to replace the inlet filters CAUTION Make sure that the entire system tubing is flushed with demineral ized water before starting the replacement of tubing In this section This section contains the following subsections Section See page 8 8 1 Replace the inlet filters 239 8 8 2 Replace the tubing and connectors 240 238 KTA start Operating Instructions 29 0270 57 AG 8 Maintenance 8 8 Replacement of tubing and filters 8 8 1 Replace the inlet filters 8 8 1 Replace the inlet filters Maintenance interval Replace the inlet filters when required for example when the visual inspection shows that the filters are clogged or damaged Required items The following items are required e inlet filters support nets Note An Inlet Filter Set containing inlet filters and support nets is included in the accessories kit Instruction Follow the instruction below to replace an inlet filter and a support net Note The inlet filters are mounted on the inlet tubing at the end that will be immersed in the buffer solution Step Action 1 Pull off the inlet filter and the support net from the inlet filter holder 7 Ny FEN a caress Ls p 3 Eas Po oo il 2 Fit the new support net and inlet filter and press the filter in position into the inlet filter holder AKTA start Operating Instructions 29 0270 57 AG 8 Maintenance 8 8 Replac
42. established by the Chinese standard SJ T11363 2006 Requirements for Concentration Limits for Certain Hazardous Sub stances in Electronic Information Products AKTA start Operating Instructions 29 0270 57 AG 27 2 Safety instructions 2 3 Emergency procedures 2 5 Emergency procedures Introduction This section describes how to do an emergency shutdown of AKTA start The section also describes the result in the event of power failure Emergency shutdown In an emergency situation Switch off power to the instrument by pressing the power switch to the O position or by disconnecting the power cord from the instrument The run is interrupted immediately WARNING Access to power switch and power cord with plug Do not block access to the power switch and power cord The power switch must always be easy to access The power cord with plug must always be easy to disconnect 28 AKTA start Operating Instructions 29 0270 57 AG 2 Safety instructions 2 3 Emergency procedures Power failure The result of a power failure depends on which unit is affected Power failure to will result in AKTA start e Therunis interrupted immediately f j The data collected up to the time of the power fail ure is available on the USB memory stick or if the system is connected to a computer UNICORN start UNICORN start on a The computer with UNICORN start installed shuts computer down On the KTA start
43. instructions below to prepare the column for a run The Column preparation procedure is initiated from the Instrument Display Step Action 1 Immerse the buffer inlet tubing in the intended buffer 2 Connect the column into the flow path For detailed instructions see Sec tion 5 5 Connect a column on page 121 3 In the AKTA start home screen tap Method run AKTA start Method run Manual run Create method Settings and service 4 In the Method run screen tap Prepare system Method run Quick start Templates User defined Prepare system l a ft AKTA start Operating Instructions 29 0270 57 AG 133 5 Prepare the system for a run 5 6 Run Prepare system methods 5 6 4 Column preparation Step Action 5 Select Column preparation and then tap Select to initiate the method For detailed instructions see Section 6 4 5 Prepare system methods on page 189 Select system method 2 Pump wash A Pump wash B L gt Washout fractionation tubing O Column preparation 1 2 Back E3 EZS 6 Set the required run parameters and then tap Run to initiate the method Run parameters Column volume v Pressure limit v Flow rate v 10 ym mi min Run volume v Result The following screen opens Column preparation in progress UV absorbance 1 0 mAU Conductivity 25 0 mS cm z Flow rate 3 0 ml min Graph Pressure 0 30 MPa Tube no 6 Edit run Hold Pause f End
44. number of collection tubes 3 A O oO 5 4 80 e 70 E ite A 60 4 a 5 z 4 j f 40 z 5 2 2 3 2 30 N 5 lt N 5 20 2 2 6 lt 10 l lo lo N N o 0 A _ _ 1 a n co wo all o per AKTA start using the Outlet valve without Frac30 A representative chromatogram depicting collection using the Outlet valve in AKTA start without Frac30 is shown below 160 AKTA start Operating Instructions 29 0270 57 AG 6 Operations from the Instrument Display 6 2 Fractionation 110 co Method Run 5 6 2013 5 32 58 PM 05 30 Method workflow_5 Result gt ollection started Hold 5 6 2013 5 35 03 PM 05 30 AutoZeroUV gt OutletValve Waste gt 1 0 5 0 0 5 1 1 5 2 2 5 3 3 5 4 45 5 5 5 6 6 5 ri 7 5 8 8 5 Note e The UV to Outlet valve volume 0 27 ml is constant for all KTA start in struments if the recommended tubing lengths and i d are used e Make sure to use recommended length and ID of PEEK tubing from UV to Outlet valve to avoid incorrect calculation of the delay volume e Make sure to update the length and ID of the PEEK tubing Settings and service System Delay volume setting in case the tubing is not of the recommended length and ID KTA start Operating Instructions 29 0270 57 AG 161 6 Operations from the Instrument Display 6 3 Perform a manual run 6 3 Performa manual run
45. of KTA start and a USB port on the computer KTA start Operating Instructions 29 0270 57 AG 81 4 Installation 4 5 Install AKTA start Step Action 3 Launch UNICORN start and connect to AKTA start For detailed instructions refer to UNICORN start 1 0 User Manual Result The instrument displays the AKTA start home screen in connected State AKTA start cy Note Make sure that the system connection is established before starting the run Always make sure to be in the AKTA start home screen connected state when trying to connect from the System Control module CAUTION KTA start is filled with denaturated alcohol 50 C2H5OH ethanol at delivery The denatured alcohol mixture can be hazardous to humans if consumed Flush out the denaturated alcohol before assembling testing or integrating AKTA start into the intended process context 82 AKTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 Prepare the system for a run About this chapter This chapter describes how to start the instrument and prepare the system for a run In this chapter This chapter contains the following sections Section See page 5 1 Flow path overview 84 5 2 Start the instrument 87 5 3 Calibrations 88 5 4 System performance 106 5 5 Connect a column 121 5 6 Run Prepare system methods 125 5 7 Sample application 135 5 8 Prepare the Fraction collector 147 5 9 Cold room operations 151
46. of modules and other module specific problems and corrective actions refer to the AKTA start Maintenance Manual Refer to the UNICORN start 1 0 User Manual for problems related to the software Note For detailed description of Warning messages refer to the AKTA start Maintenance Manual Troubleshooting procedure To troubleshoot AKTA start follow these steps Step Action 1 Follow the checklists below for system flow path and purification Z If problems remain search for solutions in Section 9 2 Basic troubleshooting on page 245 3 If problems remain after corrective actions generate a System error report see Section 3 3 4 Description of Settings and service on page 51 and contact your local GE Service Engineer System checks e Does the instrument display show the KTA start home screen AKTA start 2 Create method Settings and service e Does the ventilation of the instrument function Contact a GE Service Engineer if the fans at the bottom of the instrument stop to run AKTA start Operating Instructions 29 0270 57 AG 243 9 Troubleshooting 9 1 Introduction to troubleshooting Flow path checks Is all tubing connected correctly as shown in Section 5 1 Flow path overview on page 84 Is there leakage at any of the connections Tighten the connections if required Is any tubing bent or twisted Adjust the tubing position to make sure that the flow of liquid is smooth or replace the tub
47. result file offers the user UV curve data with fractionation marks e The result file contains 4 hours of run data e The BMP file displays the necessary legends like Product name run details UV Frac marks e Incase of shutdown or power failure the partial result will not be available However forcible ending of the run by the User results in the export of result as BMP Generate a BMP file Step Action 1 Connect a USB memory stick to the instrument 2 Start a Method run or Manual run 208 KTA start Operating Instructions 29 0270 57 AG 6 Operations from the Instrument Display 6 6 Manage methods and files 6 6 3 BMP result file Step Action 3 Tick the Save results to USB check box in order to save the results to the USB memory stick Note A BMP file is only generated when the result is saved Run parameters Column volume ml Flow rate ml min S Pressure limit MPa l rs us ES Rn 4 When the run is completed the result will be saved and a BMP file will be generated Note Do not unplug the USB memory stick when the BMP file generation is in progress 5 Transfer and connect the USB memory stick to a computer or else the image file will not be generated Open the generated BMP file view or print the re sult KTA start Result DS04 V Frac Mark nce mAU UV Absorba L Asy a 23 8 10 2 13 6 17 0 20 4 27 2 30 6 Volume ml KTA start Operating Ins
48. retin Tre E E EE 179 6 4 4 User defined MICTIOOS u 187 6 4 5 Prepdie vstenmn menoa uerius AR rere me rte eenvrtnt wrerrreyrey 189 6 5 Procrae a Ce fra U0 eee eenee ete AE 192 6 6 Manage methods and files wa eecceeccssssesssesssessssesssecssesssecssecssecssscssscsssessssessesssecssecssecsssceseesseessneesseeses 195 6 6 1 Creole COG faeces eee arate eee ee ees ee 196 6 6 2 Handling the USB memory SUCK sismiriinarisiesaninnnn 205 6 6 3 E E E AEE ENTANA VEA A ENEA 208 7 Operations from UNICORN start seesseesesseeseeserseeseeseesserserseeseeseeseessesserseeseeseese 210 PL CONTO een aE A E E E OAE 211 7 2 MEGO EOOT serre aE E EE E ence eter 213 7 3 FE UN E csc ti I ANE EA A AE E 215 CA AA a O a A E EAA EE E 216 o PONE n A O EE 217 8 1 Regular MANTENGNTCE cat ccc A 218 8 2 Cleaning before planned maintenance service ss sssssssssessssssrrrsssrresserressserresssrrrssserrrsss 221 83 Cieaning the system Tow PAUN secies ieena EEn aa E a Er 222 8 3 1 Disconnect the Colu csczscczc sates cachet RAR 225 8 3 2 Sylen 210 21 4 6 Uren ran even eto ID eae NRT TOTO RE SR 224 oi mms 10 11016 me UVTOWTEN roire E 22T 85 Cleaning the Conductivity NOW Cell sesasine eE N 229 86 Othercleanng proceduren sepenan n R E EEE 230 8 6 1 Cleaning the inlet filters eeccsecssescssessssssssessssecsssssssescssscssecssuecsssssssecsssecssecssucsssscssecsssecsueessecesses 231 8 6 2 Cleaning the instrument externally isiin 232 8 6 3 Cleaning the Fracti
49. that the Superloop is filled with liquid according to the Superloop instructions 3 Attach the Superloop to the column holder 4 Connect the tubing from the bottom of the Superloop to port 2 of the Injec tion valve AKTA start Operating Instructions 29 0270 57 AG 137 5 Prepare the system for a run 5 7 Sample application Step Action 5 Connect the tubing from the top of the Superloop to port 5 of the Injection valve 6 Make sure that a waste tubing is connected to port 4 of the Injection valve Prime the sample tubing using the Pump Follow the instructions below to prime the sample tubing using DM water Buffer before loading the sample using the Pump Step Action l Connect a 1 mm i d ETFE tubing to port I Sample of the Sample valve 2 Immerse the other end of the sample inlet tubing in the DM water Buffer container 3 From the Instrument Display select Manual Run For more details on manual run see Section 6 3 Perform a manual run on page 162 4 Tap the forward arrow to access the run parameters on screen 2 2 138 AKTA start Operating Instructions 29 0270 57 AG Step 5 Prepare the system for a run 5 7 Sample application Action Toggle to set Sample valve position to Sample so that the flow is delivered from the sample inlet Manual run Sample valve Wash valve Outlet valve Fractionation volume Tap Run to start the run End the run manually once the priming with required v
50. the Create method screen tap Delete to access the methods Select method to delete Method A Method A Method Method Cancel r e Tap a checkbox to select a method e Tap Delete to delete the method Result A Message screen that requires to confirm the action opens or To cancel the action and return to the Create method screen tap Cancel Note Multiple files can be deleted at the same time Message Are you sure you want to delete the selected method s Yes No Tap Yes to confirm the deletion of the selected files or tap No to cancel the action and return to the file list AKTA start Operating Instructions 29 0270 57 AG 6 6 2 Introduction 6 Operations from the Instrument Display 6 6 Manage methods and files 6 6 2 Handling the USB memory stick Handling the USB memory stick AKTA start provides the user with the option to store result data on a USB memory stick The USB memory stick is used to save a result BMP file and also for transferring methods between the instrument and UNICORN start The USB memory stick is also used to gen erate a System error report Note e The result files will be saved in the GE folder which is automatically created by the instrument once the USB memory stick is plugged in e At any given point of time only 10 results can be stored in the GE folder To save further results transfer the result files to another folder PC or re name th
51. the checklist in the On Site Service Health amp Safety Declaration Form or the Health amp Safety Declaration Form for Product Return or Servicing depending on whether the instrument is going to be serviced on site or returned for service respectively Copy the form you need from Section 10 4 Health and Safety Declaration Form on page 269 or print it from the PDF file available on the User Documentation CD AKTA start Operating Instructions 29 0270 57 AG 221 8 Maintenance 8 3 Cleaning the system flow path 8 3 Cleaning the system flow path Introduction Cleaning of the system flow path is performed to prevent carryover between runs con tamination in the flow path and as a routine maintenance protocol Cleaning the system flow path is usually performed by using System cleaning or Pump wash methods Note Before cleaning the system flow path remove the column from the flow path For detailed instructions see Section 8 3 1 Disconnect the column on page 223 WARNING Hazardous biological agents during run When using hazardous biological agents run the System cleaning method to flush the entire system tubing with 1M NaOH and subsequently with distilled water before service and maintenance NaOH can cause health injurious avoid spillage CAUTION Hazardous substances When using hazardous chemical and bio logical agents take all suitable protective measures such as wearing protective glasses and gloves re
52. via Loop For detailed instructions on sample applica tion refer to Section 5 7 Sample application on page 135 Vv e Set the run parameters Sample volume the volume of sample to be loaded onto the column Equilibration volume the volume of buffer A required for for equili brating the column Wash unbound volume volume of buffer needed after the sample application to wash off the unbound molecules Note The Washout unbound volume is applicable only for AC IEX techniques 198 AKTA start Operating Instructions 29 0270 57 AG 6 Operations from the Instrument Display 6 6 Manage methods and files 6 6 1 Create method Step Action gt Set parameters Elution option socratic Gradient Conc B v 10 A Elution volume v 5o yom CV Fractionation v er A ml 3 3 e Configure the run parameters for Elution Option set as Isocratic Conc B concentration of buffer B to elute the bound protein Elution volume volume needed to elute the bound protein from the column Fractionation volume volume of the fraction to be collected when the Fraction collector is enabled or Set parameters 2 Elution option Isocratic Gradient Target cone B Gradient volume CV Fractionation ml 3 3 ES E e Configure the run parameters for Elution Option set as Gradient Bound proteins are eluted with continuous change of buffer B composition to increase eluent strength over specified time Targe
53. volume of more than 1 liter each on the Buffer tray The total allowed weight on the Buffer tray is 5 kg CAUTION Avoid spillage and overflow Make sure that the waste tubing Is inserted in an appropriate waste container and secured in place 86 AKTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 2 Start the instrument 5 2 Start the instrument Switch on the instrument Follow the instruction below to start the instrument Step Action 1 Switch on the instrument by pressing the Power switch to the I position Result The instrument starts and initializes the display showing the AKTA start home screen AKTA start 2 Create method Settings and service 2 You can start using the instrument immediately All modules are pre calibrat ed from the factory KTA start Operating Instructions 29 0270 57 AG 87 5 Prepare the system for a run 5 3 Calibrations 5 3 Calibrations Introduction This section describes how to calibrate the Instrument Display Pump and Monitors In this section This section contains the following subsections Section See page 5 3 1 Calibration guide 89 5 3 2 Instrument Display calibration 90 5 3 3 Pressure sensor zero offset 92 5 3 4 Pump calibration 94 5 3 5 UV Monitor calibration 96 5 3 6 Conductivity Monitor calibration 100 88 AKTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 3 Calibrati
54. 000001 2 Delay volume Firmware update setting Ra Export system Switch valve timing report to USB A 54 Function Delay volume setting To set the delay volume The delay volume represents the volume of the liquid found in the flow path between the UV and the collection tubes Firmware update To update the system firmware Switch valve timing To set the Switch valve timing It is recommend ed to optimize the timing of switch valve Buffer valve when wavy gra dients are obtained or when fluctua tions in the step gradient are ob served during either system perfor mance tests or chromatography runs Export system report to USB Exports system error logs to a USB memory stick in a text format A system error report is useful when trying to understand a possible error that requires remote service support Note The serial number displayed on the top left of the display screen is the unique identifier for the instrument The number is also seen as instru ment ID when connecting to UNICORN start KTA start Operating Instructions 29 0270 57 AG 3 System description 3 3 Instrument Display 3 3 4 Description of Settings and service Settings and service Screen 2 The options available for the modules displayed on Settings and service Screen 2 are briefly described below Module Function Buffer valve Valve position Shows which inlet connection in the valve is open eens ka Buffer A Buf
55. 1 Check that no run is in progress 2 Switch off KTA start and disconnect the Frac30 Cable 3 Remove the collection tubes and disassemble the Bowl assembly from the Base unit e Gently move the Dispenser arm counterclockwise to the end position 1 e Push the drive assembly and hold it at the retracted position 2 e Lift and remove the Bowl assembly 3 AKTA start Operating Instructions 29 0270 57 AG 233 8 Maintenance 8 6 Other cleaning procedures 8 6 3 Cleaning the Fraction collector Step Action 4 Wash the Bowl under a water tap Use a mild detergent if required and rinse thoroughly with water 5 Wipe the surface with a damp cloth Wipe off stains using water 6 Let the Bowl assembly dry completely before re assembling 7 Re assemble the Bowl assembly on to the Base unit e Orient the Bowl to match the aligning groove and the aligning features on the Bowl holder 1 e Lower the bowl assembly on to the Base unit 2 and push the drive as sembly to allow the Bowl assembly to get in position 3 234 AKTA start Operating Instructions 29 0270 57 AG 8 Maintenance 8 6 Other cleaning procedures 8 6 3 Cleaning the Fraction collector Removing the tube holder Note The tube holder is not intended to be open by the user If required the tube holder may need to be removed from the Bowl assembly for cleaning Follow the instructions below to remove and then place back the tube holder in the Bowl assembly Ste
56. 2 Basic troubleshooting Description Possible cause Action Warning 111 UV UV Module is Mount flow cell securely and re calibrate intensity low not calibrated the UV module with the flow cell filled properly with DM water If the problem persists replace the UV module For details refer Flow cellis not to the KTA start Maintenance Manual clean Flow cell is not mounted prop erly UV LED intensi ty is below de sired level Warning 112 UV Calibration Mount the flow cell securely clean and intensity high was not per re calibrate UV module with the flow cell formed on a filled with DM water For details refer to clean flow cell the KTA start Maintenance Manual Flow cell is not mounted prop erly Warning 115 Flow cell not Mount the flow cell properly Re calibrate Flush flow celland mounted properly the UV module with flow cell filled with mount securely or is optically mis DM water aligned Warning 116 UV Flow cell is not Clean the flow cell thoroughly with DM base lining ignored clean water Re calibrate the UV module with the flow cell filled with DM water KTA start Operating Instructions 29 0270 57 AG 247 9 Troubleshooting 9 2 Basic troubleshooting Conductivity curve 248 Description Incorrect or unsta ble reading or Out of specification conductivity val ues Possible cause Conductivity cell not clean Action Clean the Conductivity cell
57. 2 Handling the USB memory stick 205 6 6 3 BMP result file 208 AKTA start Operating Instructions 29 0270 57 AG 195 6 Operations from the Instrument Display 6 6 Manage methods and files 6 6 1 Create method 6 6 1 Create method Create method menu 196 The Create method menu allows the user to create a new method edit import and delete a method from the AKTA start Instrument Display Follow the instructions below to access the Create method options Action AKTA start 2 Create method Settings and service In the KTA start home screen tap Create method Create method EJ In the Create method screen the following options are available Create to create a new method using a predefined template Edit to edit a method or change run parameters for user created methods stored on instrument USB Import to import a method developed on UNICORN start to the instru ment using a USB memory stick Delete to delete a method that is stored on the instrument KTA start Operating Instructions 29 0270 57 AG 6 Operations from the Instrument Display 6 6 Manage methods and files 6 6 1 Create method Create a method Follow the instruction below to create a method using a predefined template Step Action 1 Create method EJ In the Create method screen tap Create l Result The Select templates screen opens Select templates Affinity ac C Desalt
58. 29 0270 57 AG 2 Safety instructions 2 5 Declaration of Hazardous Substances DoHS 2 5 Declaration of Hazardous Substances DoHS AB HESJ T11364 2006 8 FERS mR AMER PERM RAK RABIES The following product pollution control information is provided according to SJ T11364 2006 Marking for Control of Pollution caused by Electronic Information Products BSS m RE Blip in Explanation of Pollution Control Label Aine WAT i SAIS I T11363 2006 E FERA Pas SVAN 205 EZR PRENABASUNA MERKATA AA MHRA AAA PmELREAWAE ABASMAFAREREMAE APEA oo RANMA RP BSARNEAR UPGKrSRSHN RR BUA 7A PRUE Pir FR BA AY R52 FB BRR DR oF PER RET E mie FAP eT oS FT PAE AY ASM RAE on PRAM ES eA RB SIR EA Ris HARARE AHRS A ELEN ASR EAR WEIR melee SRA LOTR REPENS ARTE DARE PIT AR AR AY 2 7 on BY oe EA BBR ZS a Te 52 FA A on 25 RNT PE SE A Bh I SS Sh This symbol indicates the product contains hazardous materials in excess of the limits established by the Chinese standard SJ T11363 2006 Requirements for Concentration Limits for Certain Hazardous Substances in Electronic Information Products The number in the symbol is the Environment friendly Use Period EFUP which indicates the period during which the toxic or hazardous substances or elements contained in electronic information products will not leak or mutate under normal operating con ditions so that the use of such electronic information products will not result in any severe environmental pollution any bo
59. 3 2 Instrument Display calibration 5 3 2 Instrument Display calibration Instruction Follow the instructions below to calibrate the Instrument Display Step Action i In the AKTA start home screen tap Settings and service to access the in strument modules Tap Next Next to access the 3rd screen Result The Settings and service screen will be displayed Settings and service ibe Display Conductivity 3 3 2 In the Settings and service screen tap Display to access the touch screen options Result The Display screen opens Display Touch screen f Color test calibration 90 KTA start Operating Instructions 29 0270 57 AG Step 5 Prepare the system for a run 5 3 Calibrations 5 3 2 Instrument Display calibration Action 5 5 In the Display screen tap Touch screen calibration to start the calibration Result A Message screen opens Message Touch Screen will be Calibrated now Do you want to continue Yes No Tap Yes to confirm and proceed with the calibration or Tap No to cancel the action and return to the Display screen In the four subsequent Calibration screens tap precisely on the marked circles as requested The first Calibration screen is presented as an example Calibration in progress press the marked circle shown Tap Exit when the calibration is completed Calibration done press exit Note Repeat the procedure if calibration fails AKTA sta
60. 5 9 Troubleshooting 9 3 System error report 9 5 Introduction System error report A System error report can be generated during a troubleshooting case with information about the problem The report can then be sent to GE Service Engineer for action Generate a System error report 256 Follow the instructions below to generate a System error report Note Step Make sure that a USB memory stick is connected to the instrument If the in strument cannot detect a USB memory stick the report file cannot be saved Action l In the KTA start home screen tap Settings and service to access Screen 1 see Section 3 3 4 Description of Settings and service on page 51 and then select System Settings and service Screen 1 Settings and service 5 Fraction collector Pressure sensor Pump System 1 3 Q Next Insert a USB memory stick into the USB port located on AKTA start In the System screen select Export system error report to USB System ER Serial 00000000000000000001 Delay volume Firmware update setting ie Export system Switch valve timing report to USB Back EJ Result Two files with the names INSTLOG TXT and ERRORLOG TXT are export ed to the USB memory stick KTA start Operating Instructions 29 0270 57 AG 9 Troubleshooting 9 3 System error report Step Action 3 Remove the USB stick from the USB port and connect it to a com
61. 59 41 SCS FS IT ST Scere ces avcnppeamescintatcsnecncecay nant Taasen Ee Aea ONE EEEE ain 60 4 2 Transport AK TA start and FEC 3 Oa ssctesssaain vicscisevcctivenreestanan taatvsetstpaveeironliatareeddntitomencainn 62 4 3 Unpack KTA start and Frac3o sasivussovccrrrivveseccveccacustosssicacnersasestoneebavsnennisngonedeauainwenuentitens 64 44 Accessories 610 Gl 8 G apne ne nT oan OS enii 74 OS am 6106 E E ee Renee EE enE Prev nT On OUEST 5 5 Prepare the system fOr a run ccscccsscssscssscssssccsscssscesccessecsssecssecsseessessseessseess 83 5 1 FOWA TOVE AN E EEEE T E E E 84 Se eNe eT UNE a NEA 87 e E S oei 3 9c EENE E ANEA A AON N AAA 88 53 1 Bol fo g014 61 ako 8 6 E airiran T N O A EE REENE NEN O AEAT he eran tEes 89 532 Instrument Display CONOICUOM aecenas ai i e e s E EEEE EREE 90 555 PYESSUIS sensor ZCI OOS OL csc ecsctaccecacexssscessctayctsics assists eectapeasdiaioan tary eae toeetvabnestaeesnies 92 534 PUMO oe 0 ge e emmamemen mee ein si ee ees ae entree meee mea EE A OAS 94 535 UV Montor cal oratio arses eet ee Oe A emo news 96 5 3 6 Conductivity Montor CONN OT ON sassoni Cae es iecateecteeeds staat eeeatcees 100 Manes 15 11 oe 1S 00 6 g meee me nen tet oct OR Ce re EEEa 106 541 System perlormance Method esheets acs taovecreeee iene RAE ANE 107 542 System performance Method from AKTA start u ceccssessssescssesssessssessssecssessssessssecssessssecssses 109 5 4 3 System perform
62. 8 1 Regular maintenance 8 1 Regular maintenance Introduction Regular maintenance is important for safe and trouble free operation of AKTA start The user should perform preventive maintenance on a daily weekly and monthly basis Safety precautions To avoid damage to person when performing maintenance on the instrument follow the instructions below WARNING Electrical shock hazard Do not open any covers or replace parts unless specified in the user documentation All other repairs should be done by service personnel authorized by GE WARNING Disconnect power Always switch off power to the instrument be fore replacing any component on the instrument unless stated otherwise in the user documentation WARNING Spillage Hazard Avoid spillage of fluids on the surfaces of the in strument which have cables plugs and other wirings Be careful if there is spillage of fluids on the tray while trying to remove the tray from AKTA start CAUTION Always use appropriate personal protective equipment during op eration and maintenance of AKTA start 218 AKTA start Operating Instructions 29 0270 57 AG Periodic maintenance schedule The periodic maintenance schedule provides a guide to the user to ensure proper routine maintenance of AKTA start Interval Daily Maintenance action Visually inspect the instrument for leakages in the flow path Check the Pump for leakage If there ar
63. 9 0270 57 AG 261 10 Reference information 10 1 Specifications Valves Buffer Sample Wash and Outlet Parameter Type Solenoid type switch valve No of Ports Injection valve Parameter Type 3 ports e Buffer valve and Sample valve 2 in 1 out e Wash valve and Outlet valve 1 in 2 out Rotary type manual valve Function Sample injection through Loop No of Ports Gradient formation Parameter Gradient flow rate range 6 ports 0 5 to 5 ml min Gradient composition accuracy Pressure sensor Parameter Placement of sensor 5 Conditions 5 to 95 B 1 to 5 ml min 0 8 to 2 cP and fresh pump tubing Pressure sensor is located after the Pump Range O to 0 5 MPa 5 Bar Accuracy 262 0 05 MPa AKTA start Operating Instructions 29 0270 57 AG 10 Reference information 10 1 Specifications UV Parameter Wavelength range 280 nm 3 nm Single wavelength Absorbance range 0 1 to 2 AU Linearity Within 5 up to 1 5 AU Operating pressure O to 0 5 MPa Flow cell 2 mm optical path length Total cell volume is 30 ul Conductivity Parameter Conductivity range O mS cm to 300 mS cm Resolution 1 mS cm Accuracy 5 or 2 mS cm whichever is greater Operating pressure O to 0 5 MPa Flow cell Volume 22 ul Temperature monitor 4 C to 35 C range Temperature monitor ac 10 or 5 C whichever is
64. 94 Pressure sensor Zero offset 92 Priming sample loop 140 sample tubing 138 Pump calibration 94 install tubing 76 Pump wash A instructions 126 Pump wash B instructions 128 Purpose of this document 6 R Recycling decommisioning 30 electrical components 30 general instructions 30 hazardous substances 30 procedures 30 Reference information ordering information 271 recycling 30 Regulatory compliance 10 Regulatory information international standards 10 Run parameters specifications 158 S Safety notices 8 Safety precautions emergency procedures 28 flammable liquids 17 general precautions 16 introduction 16 labels 24 25 AKTA start Operating Instructions 29 0270 57 AG personal protection 17 18 218 Sample application alternatives 135 from Pump 139 sample loop 140 Superloop 144 Sample loop connection 137 load sample 141 priming 140 Specifications 259 Starting a run final checks 153 Storage 193 Superloop connection 137 Switch valve timing settings 118 System cleaning 224 System error repor generate 256 System performance acceptance criteria 111 114 switch valve timing 117 system performance method 106 system performance method from AKTA start 109 system performance method from UNICORN start 114 7 Templates KTA start Operating Instructions 29 0270 57 AG Index Affinity AC or lon exchange IEX 180 Gel filtration Desalt ing
65. AKTA start Operating Instructions Original instructions Table of Contents Table of Contents EN cece cas tee eects sce cases ceca oc sap depette disc tsenes meteseseeenates 5 MM ADOUCIR eaten ease tea 6 1 2 HVA OT UGA userin onma sierosa anane T ANA 7 ls TRIO MOonnaUON hie aaia eee eee ee 9 1 4 Associated dOCUMENntAtiOn e sessssssssssssssssssssssssessetsttetttteeteeteeteetensssssssnsssserssdstrdtretretttreereestessssssss 12 2 Salety INSruUCTOnS sssaaa ae S 15 al Salet preca lonE sursson iae oo ne I ee EEE aN 16 e LAO AE E E AE AE A E 24 2 3 Emergency proceduras sarai E E E AE EE E EE 28 Pra REVNO DOCE OUNES cereri E AR 30 2 5 Declaration of Hazardous Substances DOHS oe ecccecsesssesssessesseesseessessesssesssecsecsssesseesseeses 31 aera E OEE TEENETE 33 31 Sy SUSI VETEN esene E A 34 5 2 N STAN E AE EE EE ON EOT E AOA TOOT 38 2o 1H GU 5 1210811 6 se ee EO EET REF EPSP 1 eU MEU eOn Sr CEES 41 5 5 4 Overview of the Instrument Display ssscsssctssstesscenscesseescrces itera tesiseindie neti nenvennnnneeee 42 3 3 2 Description OF Method TUN suisiisisereisieierisiroreissrerinieniireneist taina eetas kanani 47 3 3 3 Description of Create method sssssssesssssssirisserressssssssssssssrriresresssssssssssrrrrrrresssresssssssssren 50 3 3 4 Description of Settings and service eseccssessssssssessssessssecsssesssecsssecssessssessssecssecssueessueessecessecsseces 51 A EE O A er ee eee ee eee See eee
66. B Port I Sample Inlet tubing used when the sample is applied via the Pump Port Il Pump tubing Port III Inlet tubing from the Mixer Port I Waste Outlet tubing used when cleaning the flow path or changing the buffer by running the Pump Wash A B template Port Il Inlet tubing from the Pressure sensor Port III Outlet tubing to the Injection valve Injection valve LOAD Port 1 Ports 2 and 5 Outlet the tubing is connected to the column Inlet outlet for connecting the sample loop Port 3 Inlet for injecting the sample into the loop Port 4 Waste Outlet tubing to waste helps in washing or draining excess sample from the loop Port 6 Inlet the tubing is connected to Wash valve Outlet valve Port I Waste Outlet tubing to the waste container Port Il Inlet tubing from the Conductivity Monitor Port III Collection AKTA start Operating Instructions 29 0270 57 AG Outlet tubing to the Fraction collector 85 5 Prepare the system for a run 5 1 Flow path overview Placement of buffer bottles Buffer bottles are placed in the Buffer tray on top of the instrument as illustrated below Sample bottle or tube may be placed on the bench on the left side of the instrument A waste bottle may be placed on the bench on the right side of the instrument CAUTION Max weight on Buffer tray Do not place bottles with a
67. Base unit 2 NX me 8 8 l M ever NOTICE Never use the Dispenser arm assembly to lift or hold Frac30 To lift the module use the handle provided on the bottom plate gt Note Do not damage or break the warranty seal label during unpacking of Frac30 J NNN AKTA start Operating Instructions 29 0270 57 AG 73 74 4 Installation 4 4 Accessories package 44 Accessories package Illustration of the accessories package The illustration below shows the accessories box and the user documentation included with AKTA start at delivery AKTA start _ Lu e 113 it AKTA start Product Documentation ii if rs gt x gt z a i en D sil oe eee he iif iti SHE siS ti Gg 1s i es 5 i L s Description Unpacking Instructions System certificate Product documentation Maintenance Cue Card System Cue Card CD containing user documentation files and an unpacking video The CD includes Operating Instructions and Maintenance Manual in English and translated versions Accessories box KTA start Operating Instructions 29 0270 57 AG 4 Installation 4 5 Install AKTA start 45 Install KTA start Introduction This section describes how to install AKTA start The following actions must be performed e Install pump tubing e Connect power supply to KTA start e Connect Fr
68. Buffer Exchange 183 predefined 179 Troubleshooting basic 245 procedure 243 system error report 256 User information mes sages 253 Tubing replace tubing 240 Typographical conventions 6 U UNICORN start overview 210 Unpacking accessories package 74 USB memory stick troubleshooting 255 User information important 7 User information mes sages 253 UV flow cell cleaning 227 flow cell path length 96 UV Monitor UV LED calibration 98 W Washout fractionation tubing instructions 130 Workflow 156 281 For local office contact information visit www gelifesciences com contact GE Healthcare Bio Sciences AB Bj rkgatan 30 751 84 Uppsala Sweden www gelifesciences com AKTA GE and GE monogram are trademarks of General Electric Company AKTA HiPrep HiTrap Superloop and UNICORN are trademarks of General Electric Company or one of its subsidiaries Decon is a trademark of Decon Laboratories Ltd Deconex is a trademark of Borer Chemie AG Zuchwil Switzerland Eppendorf is a trademark of Eppendorf AG Falcon is a trademark of Becton Dickinson and Company Macintosh is a trademark of Apple Inc Marprene is a trademark of Watson Marlow Limited Microsoft and Windows are registered trademarks of Microsoft Corporation Triton is a trademark of Union Carbide Chemicals and Plastic Company Inc Tween is a trademark of Uniqema Americas LLC All other third party trademarks are the property of thei
69. CORN start 1 0 User Manual This section gives an overview of the AKTA start System Illustration of the system 34 The illustration below shows the AKTA start System with UNICORN start installed on a computer Description AKTA start instrument Frac30 Fraction collector UNICORN start software installed on a computer AKTA start Operating Instructions 29 0270 57 AG 3 System description 3 1 System overview Illustration of the instrument The illustration below shows the main parts of the instrument d Cz Description Function Instrument Display User interface for controlling the system and visualization of the runtime data Wet side The modules interconnected by tubing have the following functions e to deliver the liquid in a specified flow path and divert the flow as required e to monitor the UV absorbance and conductivity of the liquid Buffer tray Location intended for the placement of buffer bottles used during chromatography runs Power switch Connects or disconnects the power USB port To connect a USB memory stick for storage of results and transfer of files AKTA start Operating Instructions 29 0270 57 AG 35 3 System description 3 1 System overview Illustration of the Fraction collector The illustration below shows the Fraction collector Frac30 Description Function Dispenser arm Holds and positions the tubing holder for d
70. Calibration Conductivity Calibration Configuration Advanced calibration Result The Calibration screen opens Calibration 2 Oooo O M Set Conductivity EM s60 PH mS cm Calibrate Cell Temperature C A Enable Temperature Compensation E Back Note the current temperature of the calibration solution in the Conductivity flow cell as displayed in the Cell Temperature field In the Calibration screen set the theoretical conductivity value at the current temperature in the Set Conductivity field and then tap Calibrate to carry out the conductivity calibration Note e facertified conductivity standard solution is used use the supplied theo retical conductivity value corresponding to the temperature in question e Ifa manually prepared 1 00 M NaCl calibration solution is used get the conductivity value at the current temperature from the graph for conduc tivity of the 1 00 M NaCl as a function of temperature presented below AKTA start Operating Instructions 29 0270 57 AG 103 5 Prepare the system for a run 5 3 Calibrations 5 3 6 Conductivity Monitor calibration Graph for Conductivity value The graph below shows the Conductivity value at the current temperature when 1 00 M NaCl calibration solution is used Conductivity of 1 00 M NaCl at 20 30 C
71. ICORN start Instructions Follow the instructions below to initiate the System performance method from UNICORN Start Step Action 1 Start the test from UNICORN start System control System Performance Test and Report Select method based on the Fraction collector configuration e Performance method with Frac when the Fraction collector is enabled e Performance method without Frac when the Fraction collector is disabled Read the method notes before starting the run Make a note of the result file location Run the System performance method The test report states whether the System performance method passed or failed Manually verify Pressure limits Fractionation collection volumes gradient levels and all connections for leakage during the test using the Acceptance criteria presented below Acceptance criteria Activity Approved interval 1 ml min 0 B flow through Back pressure lt 0 05 MPa Outlet valve waste position Repeat UV Auto zero 5 ml min Back pressure 0 06 to 0 2 MPa Start gradient 0 to 100 B in 10 minutes start fractiona tion collection 114 AKTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 4 System performance 5 4 3 System performance method from UNICORN start Activity Approved interval 1 End fractionation Weigh fraction no 0 8to1 2g 2 3 and 4 Max difference 0 19 between fractions End collect
72. Injection valve acknowledge the message by tapping Continue 146 AKTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 8 Prepare the Fraction collector 5 8 Prepare the Fraction collector Prepare the Fraction collector Fractions are collected in tubes using the Fraction collector Follow the instructions below to prepare the Fraction collector if the Fraction collector is going to be used during a run The following types of tubes can be placed in the tube holder of the Bowl assembly e Eppendorf tubes 1 5 ml or 2 ml e 5mltubes 12 x 75 mm e Centrifuge tubes 10 to 12 ml e Falcon tubes 15 ml NOTICE The Fraction collector should be connected or disconnected from the instrument only when KTA start is switched off Note Make sure that the Fraction collector is properly installed See Connect Frac30 to AKTA start on page 78 Step Action 1 Insert a sufficient number of collection tubes in to the Bowl assembly Note All the tubes must be of the same length and diameter and there should be no empty spaces in the sequence 2 Connect a 0 75 mm i d PEEK tubing to the Outlet valve port III Collection Note The tubing must be about 50 cm long to ensure proper placement of the Fraction collector and free movement of the dispenser arm 3 Loosen the nut of the tubing holder and insert the outlet tubing into the tubing holder and tighten the nut Note
73. M NaCl e Sample valve inlet immersed in sample 1 0 acetone 1 0 M NaCl e Make sure that 1 ml Sample loop is filled with sample 1 0 acetone 1 0 M NaCl AKTA start Operating Instructions 29 0270 57 AG 107 5 Prepare the system for a run 5 4 System performance 5 4 1 System performance method 108 Make sure that 2 m of 0 5 mm ID tubing is connected to the Outlet valve at Waste position When performing System performance method without Fraction collector make sure the Outlet valve fractionation tubing is inserted into a pre weighed beaker When performing System performance method with Fraction collector make sure the Outlet valve fractionation tubing is connected to Fraction collector with at least 5 pre weighed tubes Make sure that the system is prefilled with DM water Make sure to take note of all the required observations by recording parameters while the System performance method is in progress Enter the observed values in the System report template presented in Section 11 1 System Performance Report on page 276 AKTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 4 System performance 5 4 2 System performance method from AKTA start 5 4 2 System performance method from AKTA start Instruction Follow the instructions below to initiate the System performance method from the In strument Display Note Step Insert a USB memory stick to save the results Action 1
74. Prepare the system for a run 5 3 Calibrations 5 3 6 Conductivity Monitor calibration Action In the Configuration screen set the Cell Constant value in the Set Cell Const field for the new Conductivity flow cell by using the up down arrows Tap Save to save the new value Note The Cell Constant value is provided on the packaging of the Conductivity flow cell If a Cell constant value is not available calibrate the Conductivity flow cell as explained in Calibrate the Conductivity flow cell on page 103 Set the Reference Temperature Note For the system to recalculate the measured conductivity provide the reference temperature in the temperature field Set Ref Temp Make sure that the checkbox Enable Temperature Compensation in the Conductivity Calibration screen is checked Follow the instructions below to set the reference temperature Step Action 1 In the Conductivity Configuration screen set the Reference Temperature Set Ref Temp value in the range 4 C to 35 C Configuration 3 Set Cell Const 5 43o AN ijem Set Ref Temp v 200 ym C Tap Save to save the new reference temperature value KTA start Operating Instructions 29 0270 57 AG 101 5 Prepare the system for a run 5 3 Calibrations 5 3 6 Conductivity Monitor calibration Calibrate the temperature sensor 102 Follow the instructions below to calibrate the temperature sensor Step Action
75. Prepare the system for a run 5 5 Connect a column 5 5 Connect a column Introduction This section describes how to connect a column to KTA start Different types of column holders are available for the columns that are to be used with the instrument See Chapter 10 Reference information on page 258 for selecting an appropriate column holder The column is connected in the flow path between the Injection valve and UV Monitor as shown in the illustration of the flow path in Section 5 1 Flow path overview on page 84 Column placement Depending on column dimension choose the appropriate location on the instrument to place the column Column holder rails are located on the front and on the right side of the instrument as shown in the image below e Front side of the instrument for small columns e g HiTrap columns e Right side of the instrument for large columns e g column length 60 cm AKTA start Operating Instructions 29 0270 57 AG 121 5 Prepare the system for a run 5 5 Connect a column Connect a column Follow the instructions below to connect a column to the instrument Step Action 1 Attach an appropriate column holder to the column holder rail on the instru ment CONDUCTIVITY TION VALVE been Sangh n 2 Remove the column stoppers mount the column on the union connector if the column type requires a union IW wdEJLIH 122 KTA start Operating Instructions 29 0270 57 AG 5
76. Replace the pump tubing See Sec tion 8 8 2 Replace the tubing and connec tors on page 240 Pump tubing is not fixed inside the pump hood Fix the pump tubing inside the pump hood and start the run Failure of compo nent in Main board Troubleshoot the Pump and if the prob lem still persists contact a GE Service Engineer Pump Description Erratic flow Inaccurate flow rate No flow 250 KTA start Operating Instructions 29 0270 57 AG Mixer Description Leakage Description Leakage Fraction collector KTA start Operating Instructions 29 0270 57 AG Description The fractions col lected fall outside the collection tubes Possible cause Tubing connection Possible cause Tubing connection Possible cause The Bowl Assembly improperly fitted on the Base unit 9 Troubleshooting 9 2 Basic troubleshooting Action Check the tubing connections Re tighten or replace if necessary Action Check the tubing connections Re tighten or replace if necessary Action Make sure that the Bowl is fitted properly on the base not tilted Dispenser arm not in correct position Check that the dispenser arm is in the dispensing position Notice the alignment of the markings on the dispenser arm Drive sleeve is worn out and slip page occurs Replace the drive sleeve Refer to the KTA start Maintenance Manual Fraction collector is not homing at
77. See Sec tion 8 5 Cleaning the Conductivity flow cell on page 229 Cable improperly connected to the Check that the Conductivity module ca ble is connected properly to the connec Conductivity Mon tor behind the flow cell itor Pump malfunction Check that the Pump functions properly See Pump troubleshooting Temperature sen sor not calibrated Non equilibrated column Calibrate the temperature sensor See Section 5 3 Calibrations on page 88 Check that the column is equilibrated If necessary clean the column Buffer valve mal function The 0 5 mm ID 195 mm PEEK tubing is not connected between the Con ductivity Monitor and the Outlet valve Check the operation of the Buffer valve Refer to the KTA start Maintenance Manual Connect 0 5 mm ID PEEK tubing See Section 8 8 2 Replace the tubing and connectors on page 240 Failure of compo nent in Main board Contact a GE Service Engineer Baseline drift or noisy signal Air trapped in the conductivity flow cell Remove the air trapped in the flow cell by flushing the cell with DM water or buffer If persistent clean the Conductiv ity flow cell See Section 8 5 Cleaning the Conductivity flow cell on page 229 Non equilibrated column Check that the column is equilibrated If necessary clean the column Pump malfunction Check that the pump functions properly See Pump troubleshooting
78. Select method to edit screen tap a radio button to select a user method e Tap Edit to start editing the run parameters for the selected method 3 Set parameters Flow rate Zam ml min n Pressure limit v ym MPa Set parameters Samplefrom Pump Loop r Sample volume gt Equilibration M 00 io 150 Wash unbound v Set parameters 2 Elution option socratic Gradient gj ome a gt B lt Elution volume CV Fractionation v 20 yom mil 3 3 AKTA start Operating Instructions 29 0270 57 AG 201 6 Operations from the Instrument Display 6 6 Manage methods and files 6 6 1 Create method Step Action e Select Save Method as field if you want to set a method name The file name can be edited by setting the digits in the range 00 to 99 Note Provide a unique method name for example ACO2 DSO5 The created method will be saved under the User defined methods menu USB imported methods which were created using UNICORN start cannot be edited from the instrument Use UNICORN start to edit those methods Message A Are you sure you want to save the method Yes No Tap Yes to confirm the saving of the method or Tap No to cancel the action and return to set the run parameters Import a method Follow the instruction below to import a method stored on a USB memory stick Note Make sure that the USB memory stick containing the user defined methods is con
79. System performance 5 4 4 Switch valve timing Step Action fa Tap System Result The System screen opens System 2 Serial 00000000000000000001 its setting i om Export system eok j ca 3 Tap Switch valve timing Result The Switch valve timing screen opens 4 Select Switch valve timing B to change the Switch valve timing to 5 sec Switch valve timing Switch valve timing A Default Switch valve timing B Advanced timing 5 Tap Save to save the changed timing 6 If wavy gradients persist or if fluctuations on step gradient levels are large select Advanced timing to set switch valve timing other than Switch valve timing A and Switch valve timing B Switch valve timing gt Switch valve timing A Default Switch valve timing B Advanced timing Switch valve time Sec a Lo Note It is recommended to use the Advanced timing if the options for Switch valve timing A or Switch valve timing B have not yielded satisfying results i Set Switch valve time in the range of 3 0 to 5 0 sec AKTA start Operating Instructions 29 0270 57 AG 119 5 Prepare the system for a run 5 4 System performance 5 4 4 Switch valve timing Step Action 8 Tap Save to save the optimized timing 9 Perform the System performance method or 50 gradient runs until satis factory results are obtained 120 AKTA start Operating Instructions 29 0270 57 AG 5
80. To configure the new UV monitor and flow cell Note The C amb and C drft values should not be altered by the user UV Config uration is performed by a GE Service Engineer only Display Display Display Touch screen calibration Diagnostics Back E a Color test GJ Touch screen calibration To cali brate the touch screen Color test To perform a diagnostics test for the colors displayed on the touch screen Diagnostics To perform a diagnos tics test to check the backlight per formance of the display Log book To read the number of hours the instrument display has been in use AKTA start Operating Instructions 29 0270 57 AG 57 3 System description 3 3 Instrument Display 3 3 4 Description of Settings and service Module Function Conductivity Calibration To calibrate the Conduc tivity flow cell and the temperature sensor nere o Configuration To set a new cell Calibration Configuration constant value or reference temper Conductivity PEE atu re value usually after the Conduc soles tivity flow cell replacement Advanced calibration To calibrate the temperature of the Conductivity flow cell Note Calibration of the Sine Wave Gener ator should be performed by a GE Service Engineer only after the Main board has been replaced 58 AKTA start Operating Instructions 29 0270 57 AG 4 Installation 4 Installation About this chapter
81. absorbance 1 0 mAU Conductivity 25 0 mS cm Flow rate 3 0 ml min Graph Pressure 0 30 MPa Tube no 6 Edit run End While the System cleaning run is in progress wait for the run to complete Note If required the System cleaning run can be ended before it completes by tapping End i Run ended UV absorbance 1 0 mAU Conductivity 25 0 mS cm z Flow rate 0 0 ml min Graph Pressure 0 30 MPa Tube no 6 Exit Tap Exit to close the System cleaning screen when the System cleaning run is completed AKTA start Operating Instructions 29 0270 57 AG 191 6 Operations from the Instrument Display 6 5 Procedures after a run 6 5 Procedures after a run Introduction This section briefly describes e How to evaluate a recorded result e How to clean the instrument after a run e How to prepare the system for storage if the instrument is not going to be used for a period of time The instrument and the columns should be cleaned between the runs This will prevent for example carryover and cross contamination among samples protein precipitation and clogging of the column or flow path For further details on cleaning and maintenance procedures see Chapter 8 Maintenance on page 217 Evaluate a run After a chromatographic run the result stored on a USB memory stick can be transferred to UNICORN start where it can be viewed and evaluated The result holds a complete record of the run including method sys
82. ac30 to AKTA start e Connect KTA start to the UNICORN start computer WARNING Only use grounded power cords delivered or approved by GE WARNING Supply voltage Make sure that the supply voltage at the wall outlet corresponds to the marking on the instrument before con necting the power cord CAUTION Protective ground AKTA start must always be connected to a grounded power outlet AKTA start Operating Instructions 29 0270 57 AG 75 4 Installation 4 5 Install KTA start Install pump tubing Follow the instructions below to install the pump tubing Step Action 1 Open the top cover until it is fully open Z Place the tubing between the rollers and the track press against the pump head inner wall Note Make sure that the pump tubing is not twisted or stretched against the rollers 76 AKTA start Operating Instructions 29 0270 57 AG 4 Installation 4 5 Install AKTA start Step Action 3 Lower the top cover until it clicks into its fully closed position The track closes automatically and the tubing is stretched correctly as the track closes PUMP Connect power to AKTA start Follow the instructions below to connect power to AKTA start Step Action il Select the correct power cord to be used KTA start is delivered with 2 alter native power cords e Power cord with US plug 2 m e Power cord with EU plug 2 m Note Discard the unused
83. al Clean the UV flow cell every six months or when required A clean flow cell is essential for the acceptable performance of the UV Monitor NOTICE Keep UV flow cell clean Do not allow solutions containing dissolved salts proteins or other solid solutes to dry out in the UV flow cell Do not allow particles to enter the UV flow cell as damage to the flow cell may occur Required solutions The following solutions are required e Cleaning solution 10 surfactant detergent solution such as Decon 90 Deconex 11 RBS 25 1M HCl or 1M NaOH e DM water Note e tis recommended to use 10 surfactant detergent solution for cleaning the UV flow cell e Heat the 10 surfactant detergent solution to 40 C to increase the cleaning efficiency e f NaOH is used perform cleaning at 1ml min and reduce the hold time to 5 min in step 3 of the method mentioned below e NaOH should not be left in the flow cell for more than 20 minutes and proper care should be taken to remove the NaOH completely from the flow cell Cleaning the UV flow cell in place Follow the instructions below to clean the UV flow cell Note Before cleaning the UV flow cell remove the column from the flow path and re connect the flow path See Section 8 3 1 Disconnect the column on page 223 Step Action 1 Immerse the inlet tubing in cleaning solution AKTA start Operating Instructions 29 0270 57 AG 227 8 Maintenance 8 4 C
84. ample injection in the Sample from field to be ap plied via Pump or via Loop For detailed instructions on sample applica tion refer to Section 5 7 Sample application on page 135 AKTA start Operating Instructions 29 0270 57 AG 181 6 Operations from the Instrument Display 6 4 Perform a method run 6 4 3 Templates Step Action e Set the run parameters Sample volume the volume of sample to be loaded onto the column Equilibration volume the volume of buffer A required for for equili brating the column Wash unbound volume volume of buffer needed after the sample application to wash off the unbound molecules Note For AC IEX methods when loading sample through loop it is advisable to empty the loop with 3 times the loop volume to achieve good sample recovery Run parameters Elution option socratic Gradient r ConcB v 100 A lt i Elution volume v 5o ym CV Fractionation volume e Configure the run parameters for Elution Option set as Isocratic Conc B concentration of buffer B to elute the bound protein Elution volume volume needed to elute the bound protein from the column Fractionation volume volume of the fraction to be collected when the Fraction collector is enabled or Run parameters Elution option isocratic Gradient lt Gradient volume CV om BM B volume 182 KTA start Operating Instructions 29 0270 57 AG 6 Operations from the Inst
85. ance method from UNICORN start sssrinin 114 5 4 4 T OE a EE S E RENEE eee 117 a E 0 610 0 aa e E EE E E E EN 121 2 AKTA start Operating Instructions 29 0270 57 AG Table of Contents 56 R nPrepare svetem MCT OCS sepsis cc caceers ct cottetcenscerccestaveuehchesebied E ERE 125 5 6 1 PUMP WOSR A sissitoiminnan stata tide RER 126 562 POTTO E a E E EAEE ERE 128 5 6 3 Washout fractionation tUbING sisccscesacstsrsteesesnasensastsssteaveenspovsctvaienavthewetpabisettasdienvtentoernestee 130 5 6 4 Column preparo UON sorserien i n E SA 133 5 Samoe dppicalon senesinin E ER 135 ge Prepare iNe Frac on CONECTO sennae EES 147 r 00 31 6619 902 9 raO ea E N A 151 TAO A Ci ereer EE EE E E EA A OE AEEA 153 6 Operations from the Instrument Display seesesssesseseesseeseeserseeseeseesseeseeseeseeseesee 155 GL IAOU O a EE A oectetoseesteessio seaceeetes 156 6 2 Eran 10 6 M tee ener ert E E O A EE E 159 6 3 Perform a manual run es csnese inne exestces sevens tant tes acess ceecssee anne sovestuscosanadhauesonct aeeeieceenioeseaes 162 6 5 1 PNM TY EEN Sa eee ep act ee 163 6 3 2 M nit r and control the TUN ce secseccrecs teeter aerate eaten E eae tees 165 6 4 Performa method TUN anne annem nn nS totes OO avinen uonane eet nv ov PCE 170 6 4 1 Selecta 60 16810 00 0 lt a en ek ee ee re ee er en eee een meee eee nee ee ees 171 6 4 2 OLE os 6s 0 5 E E mem nT A TORE NR Ow IO Tena Teer ne Tee eT a 173 6 4 3 EO a ter perv reer trertete tin
86. ater before service and maintenance e Setting Check that the correct outlet size settings are used Make sure that tubing and fittings are properly connected and secured Make sure that the pressure limit settings are correct before starting the run 20 AKTA start Operating Instructions 29 0270 57 AG 2 Safety instructions 2 1 Safety precautions CAUTION To avoid personal injury when operating AKTA start follow the in structions below Max weight on Buffer tray Do not place containers with a volume of more than 1 liter each on the Buffer tray The maxi mum allowed weight on the Buffer tray is 5 kg Large spillage Switch off AKTA start and unplug the power cord if large spillage occurs NOTICE To avoid damage to KTA start or other equipment when operating the instrument follow the instructions below Keep UV flow cell clean Do not allow solutions containing dissolved salts proteins or other solid solutes to dry out in the flow cell Do not allow particles to enter the flow cell as damage to the flow cell may occur Prefill UV flow cell Make sure that the UV flow cell is prefilled with liquid before starting the system Avoid condensation If KTA start is kept in a cold room cold cabinet or similar keep the instrument switched on in order to avoid condensation Avoid overheating If KTA start is kept in a cold cabinet and the cold cabinet is switched off make sure to switch o
87. aturated alcohol 50 C2H5OH ethanol at delivery The denatured alcohol mixture can be hazardous to humans if consumed Flush out the denaturated alcohol before assembling testing or integrating AKTA start into the intended process context Personal protection WARNING To avoid hazardous situations when working with AKTA start take the following measures for personal protection Spread of biological agents The operator has to take all necessary actions to avoid spreading hazardous biological agents in the vicinity of the equipment The facility should comply with the na tional code of practice for biosafety AKTA start Operating Instructions 29 0270 57 AG 17 2 Safety instructions 2 1 Safety precautions CAUTION To avoid hazardous situations when working with AKTA start take the following measures for personal protection e Always use appropriate personal protective equipment during operation and maintenance of AKTA start e Spillage hazard When using KTA start use personal protective equipment like goggles lab coat protective shoes and gloves to avoid any circumstances of spillage e Cutinjuries The tubing cutter is very sharp and must be han dled with care to avoid injuries e Hazardous substances When using hazardous chemical and biological agents take all suitable protective measures such as wearing protective glasses and gloves resistant to the sub stances used Follow local and
88. automatically search the Light strength value to get a minimum Signal response of 2500 mV e If the Signal value is above 2500 mV tap Save If the Signal value is below 2500 mV use the arrows to increase the Light strength until the Signal is above 2500 mV T Tap Save to store the calibrated Light strength value KTA start Operating Instructions 29 0270 57 AG 99 5 Prepare the system for a run 5 3 Calibrations 5 3 6 Conductivity Monitor calibration 5 3 6 Conductivity Monitor calibration Set the Cell constant value If the flow cell was replaced a new Cell constant value has to be set Follow the instruc tions below to set the Cell constant value Step Action 1 In the AKTA start home screen tap Settings and service to access the in strument modules then tap Next Next Result The Settings and service screen 3 opens Settings and service Ca ae Display Conductivity Back tat E J 2 In the Settings and service screen tap Conductivity to access the Conduc tivity Monitor options Result The Conductivity screen opens Conductivity Calibration Configuration Advanced calibration 3 In the Conductivity screen tap Configuration to access the configuration options Result The Configuration screen opens Configuration 2 Set Cell Const S 430 O A ijem Set Ref Temp v 20 A C Back E J 100 KTA start Operating Instructions 29 0270 57 AG Step 5
89. ave been removed from the area to provide access Provide explanation for any No answers here Equipment type Product No Serial No hereby confirm that the equipment specified above has been cleaned to remove any hazardous substances and that the area has been made safe and accessible Name Company or institution rae or Date YYYY MM DD Signed GE and GE monogram are trademarks of General Electric Company GE Healthcare Bio Sciences Corp 800 Centennial Avenue P O Box 1327 Piscataway NJ 08855 1327 2010 14 General Electric Company All rights reserved First published April 2010 DOC1149542 28 9800 26 AC 05 2014 AKTA start Operating Instructions 29 0270 57 AG 269 10 Reference information 10 4 Health and Safety Declaration Form Product return or servicing Health amp Safety Declaration Form for Product Return or Servicing Return authorization and or number Service Ticket Request To make sure the mutual protection and safety of GE personnel our customers transportation personnel and our environment all equipment must be clean and free of any hazardous contaminants before shipping to GE To avoid delays in the processing of your equipment please complete this checklist and include it with your return 1 Please note that items will NOT be accepted for servicing or return without this form 2 Equipment which is not sufficiently cleaned prior t
90. ays the method templates for chromatography techniques avail Lec i able with KTA start iii For a description of the templates C Desalting buffer exchange DS see Section 6 4 3 Templates on O lon exchange IEX page 179 T Gel filtration GF Back ED EZS User defined Displays the methods created by the user Select method For detailed instructions on creating om and managing user defined meth 0506 ods see Section 6 6 Manage methods and files on page 195 Back Select Prepare system 1 2 Displays the predefined system methods that are used to clean the Select system method system flow path ee For detailed instructions on running O Pump wash B Fi system methods see Section 6 4 5 gt Washout fractionation tubing Prepare system methods on page 189 Column preparation 1 2 Back EZS 48 KTA start Operating Instructions 29 0270 57 AG 3 System description 3 3 Instrument Display 3 3 2 Description of Method run Method Description Prepare system 2 2 Select system method System cleaning O System performance method AKTA start Operating Instructions 29 0270 57 AG 49 3 System description 3 3 Instrument Display 3 3 3 Description of Create method 3 3 3 Description of Create method Create method options The display option Create method allows the user to create new methods edit or delete existing user methods and also import methods
91. cription The Instrument display does not show anything Possible cause No power Action Check that the Power cord is connected and that the Power Switch is switched on The Instrument display is dam aged Contact a GE Service Engineer Communication problem no sig nals to the display Contact a GE Service Engineer Issues with the touch response Calibration may have gone bad Re calibrate the Touch screen For details refer to the KTA start Maintenance Manual The Display color is not proper Display malfunc tion Display inter face is cable loose Perform a Display Color test refer to KTA start Maintenance Manual If there is a pattern mismatch contact a GE Ser vice Engineer The Display Back light is not working AKTA start Operating Instructions 29 0270 57 AG Display malfunc tion Display inter face is cable loose Perform a Display Diagnostic Backlight test refer to AKTA start Maintenance Manual Check whether the Backlight is changing or not If it is not changing contact a GE Service Engineer 245 9 Troubleshooting 9 2 Basic troubleshooting UV curve 246 Description Noisy UV signal Signal drift or insta bility Possible cause If UV noise level is above 10 mAU air bubbles are trapped in the flow cell Action Remove the air bubbles trapped in the UV flow cell by flushing the cell with DM water or buffer If
92. de no 29 0520 87 Packaging Kit for Frac30 KTA start Operating Instructions 29 0270 57 AG 90557 03 213 10 Reference information 10 5 Ordering information Service tools Torx driver T10 29 0031 71 Torx driver T20 28 9513 03 Flat screwdriver 274 56 4656 00 AKTA start Operating Instructions 29 0270 57 AG 11 Appendix 11 Appendix About this chapter This appendix presents a template for a Report of the System performance method The Report has to be filled in with observations collected during the System performance method which is performed either from AKTA start or from UNICORN start In this chapter This chapter contains the following sections Section See page 11 1 System Performance Report 276 AKTA start Operating Instructions 29 0270 57 AG 215 11 Appendix 11 1 System Performance Report 11 1 System Performance Report Test performed from AKTA start Time min Activity Check Approved interval Observations 0 Pump wash Wash valve posi Mobile phase out tion through Waste 1 1 ml min 0 B flow Back pressure lt 0 05 MPa through Outlet valve Waste position 2 Repeat UV Auto zero 3 5 ml min Back pressure 0 06 to 0 2 MPa UV level 10 mAU Conductivity lev 1 mS cm el 4 1 ml min Sample valve Max UV level 300 to 380 mAU Sample position Max Conductivi 65 to 95 mS cm ty
93. dily injury or damage to any assets The unit of the period is Year In order to maintain the declared EFUP the product shall be operated normally ac cording to the instructions and environmental conditions as defined in the product manual and periodic maintenance schedules specified in Product Maintenance Pro cedures shall be followed strictly Consumables or certain parts may have their own label with an EFUP value less than the product Periodic replacement of those consumables or parts to maintain the declared EFUP shall be done in accordance with the Product Maintenance Procedures This product must not be disposed of as unsorted municipal waste and must be collected separately and handled properly after decommissioning AKTA start Operating Instructions 29 0270 57 AG 31 2 Safety instructions 2 5 Declaration of Hazardous Substances DoHS BeA SWART ANAMRES Name and Concentration of Hazardous Substances Pras ASWANTtANEhMReS Table of Hazardous Substances Name and Concentration bE E ABA SWAN Component name Hazardous substance R 7K tf KER Pb Hg Cd Cr6 29 0220 94 29 0230 51 0 RRBZABAS VAEABAM BSS A SB ESI T11363 2006 EEI E AY PR BE RAR x RRBZABAS WASYERREHR HAMS SY SB BiB wSI 711363 2006 EM TERY PRB ESR We Ze PU BR HEA AC 7h AY PT ER SAY Be ES 0 Indicates that this toxic or hazardous substance contained in all of the homogeneous ma terials for this part is below the limit re
94. ding to the column instructions If the column is not going to be used for a couple of days or longer perform the following e Fill the column with the storage solution recommended in the column data sheet e Detach the column from the instrument and store it according to the column recom mendation System storage If the instrument is not going to be used for a period of time fill the system and the inlets with storage solution DM water or 20 ethanol For detailed instructions see Section 8 7 Storage of the instrument on page 236 AKTA start Operating Instructions 29 0270 57 AG 193 6 Operations from the Instrument Display 6 5 Procedures after a run Power off the instrument Switch off the instrument by turning the Power switch to the O position Note When the instrument is switched off or not in use make sure to open the pump hood and release the pump tubing from the pump head 194 AKTA start Operating Instructions 29 0270 57 AG 6 Operations from the Instrument Display 6 6 Manage methods and files 6 6 Manage methods and files Introduction This section describes how to create edit import and delete methods on AKTA start For information about how to create a method using the UNICORN start see Chapter 7 Operations from UNICORN start on page 210 or the UNICORN start 1 0 User Manual In this section This section contains the following subsections Section See page 6 6 1 Create method 196 6 6
95. e Fraction mAU Injection Run Log 600 400 200 0 200 400 T 7 T 7 min 0 0 05 0 1 0 15 0 2 0 25 0 3 0 35 0 4 0 45 0 5 0 55 0 6 0 65 0 7 0 75 0 8 0 85 0 9 0 95 Process Picture x ee z Ready 0 00 ms Cm Outlet valve Buffer valve n B Accumulated Time 0 min Accumulated Volume o ml ae Se E Column i 0 00 mAU e 2 Injection valve Wash valve g 0 00 B g F m l Tube number 0 50 ml min 0 00 MPa Ready Block No watch Connection Connected in control Controlled by Default HCE 21CK5R1 Main features The main features of the System Control module are listed below e A flow scheme representing the real time flow path with indications of the different modules in wet side of the instrument Current run status of the system is displayed e The ability to control the instrument via simple clicks on the flow path for example to turn the valves set flow rates change B concentrations and start stop fractiona tion AKTA start Operating Instructions 29 0270 57 AG 211 7 Operations from UNICORN start 7 1 System Control A real time chromatogram depicting the complete run with curves including UV conductivity system flow gradient concentrations fraction marks run logs and pressure The ability to perform manual and method runs Ability to run predefined methods like Quick start Templates and Prepare system methods Ability to perform System performance
96. e GE folder Store result on a USB memory stick Follow the instructions below to store results generated in KTA start on a USB memory stick Step Action il Connect a USB memory stick to the instrument via the USB port 2 Start a Manual run or Method run 3 In the Run parameters screen tick the Save results to USB check box in order to save the generated results on the USB memory stick When the run is complete the results are saved in a GE folder 4 A BMP result file is also generated Hence the chromatogram can be viewed without the use of UNICORN start 5 Export the results to UNICORN start to view the chromatogram and evaluate Note Make sure that the USB memory stick is not removed during the run For more details see Do s and Dont s while handling the USB memory stick on page 205 Do s and Dont s while handling the USB memory stick e Make sure that the USB memory stick is inserted completely into the instrument AKTA start Operating Instructions 29 0270 57 AG 205 6 Operations from the Instrument Display 6 6 Manage methods and files 6 6 2 Handling the USB memory stick Max supported USB memory stick is 32 GB Minimum 1 GB free space is required to execute read write operations Only FAT32 file system is supported and this needs to be taken into account while formatting the memory stick Only unplug the USB memory stick when the instrument display is in the Home screen It is preferred to
97. e Instrument Display Button Name Description Increment up The value in the text field can be increased or de mo arrow creased by tapping up or down arrow Decrement Tapping the value opens the numpad and a new down arrow value can be typed in Note After typing in a value on the numpad tap OK to confirm the new value Next Opens the next screen Next i Back Returns to the previous screen Back a Run Starts a run Pause Pauses the ongoing run by stopping the Pump The flow rate settings and the gradient values are retained Continue Continues a run that has been paused a Hold Holds an ongoing run with current flow rate valve Hold positions and with set B concentration The gradient is held at the value displayed es Resume Resumes a run that is put on hold Resume Eo Edit run Opens a new screen for editing the current run Edit run parameters 44 KTA start Operating Instructions 29 0270 57 AG 3 System description 3 3 Instrument Display 3 3 1 Overview of the Instrument Display Button Description Execute Executes the edited run parameters during a run Confirms a selection or an action Cancel Cancels a selection or an action Terminates the ongoing run always followed by a screen that requires to confirm the action When a run is completed it closes the application and returns to the AKTA start home screen Graph icon Ope
98. e out the document placed at the top of the package and read the Un packing Instructions 3 Holding the red strap lift the fraction collector out of the delivery box Place the Fraction collector on the laboratory bench tice ROCIO Ece KTA start Operating Instructions 29 0270 57 AG 69 4 Installation 4 3 Unpack KTA start and Frac30 Step Action 4 Open the strap lock and remove the strap 70 AKTA start Operating Instructions 29 0270 57 AG 4 Installation 4 3 Unpack KTA start and Frac30 Step Action 6 Remove the foam cushion from the bottom of the fraction collector by carefully lifting the Fraction collector 7 Remove the plastic bag AKTA start Operating Instructions 29 0270 57 AG 71 72 4 Installation 4 3 Unpack KTA start and Frac30 Step Action 8 Remove the Bowl assembly from the Base unit e Gently move the Dispenser arm counterclockwise to the end position e Push the Drive assembly and hold it at the retracted position At the same time lift the Bowl assembly 9 Remove the foam cushion located on the Base unit AKTA start Operating Instructions 29 0270 57 AG 4 Installation 4 3 Unpack KTA start and Frac30 Step Action 10 Re assemble the Bowl assembly on to the Base unit Orient the Bowl to match the aligning groove and the aligning features located on the bowl holder e Slightly push the Drive assembly laterally and lower the Bowl assembly onto the
99. e return arrow to return to the Run view screen Edit the run Follow the instruction below to edit the run parameters of an ongoing run Step Action 1 Run view UV absorbance 1 0 mAU Conductivity 25 0 mS cm 3 Flow rate 3 0 ml min Graph Pressure 0 30 MPa Tube no 6 Edit run End In the Run view screen tap Edit run to access the run parameters of the ongoing run 166 AKTA start Operating Instructions 29 0270 57 AG 6 Operations from the Instrument Display 6 3 Perform a manual run 6 3 2 Monitor and control the run Step Action Edit run Conc B v ym 2 Flow rate v 30 Zam mlifmin r gt Fractionation Fractionation volume Coone E e Edit the run parameters Conc B buffer B concentration Flow rate flow rate ml min Fractionation volume fractionation volume ml Toggle Fractionation between Start and Stop to start or stop the fractionation Use up down arrows to adjust the values or use the numpad to type in the values If no other parameters need to be set tap Execute to implement the changes To ignore the changes tap Cancel e Tap the forward arrow to access additional run parameters to edit Note Before starting the run set the status of the Fraction collector to enabled or disabled as required Edit run Sample valve Buffer Wash valve Waste Outlet valve Setevent mark Autozero UV 2 2 Cancel Execute
100. e signs of liquid leaking from the Pump check the in tegrity of the pump tubing and the tubing con nections Note Make sure that the pump tubing is not left inside the Pump when it is not in use 8 Maintenance 8 1 Regular maintenance See section Clean the column and the system flow path af ter use and leave the system filled with DM water Note If the instrument is not going to be used for a few days prepare the system for storage Section 6 5 Procedures after a run on page 192 Section 8 3 Cleaning the system flow path on page 222 Weekly or when required Calibrate the Pump Visually inspect the inlet filters and clean them if necessary Section 5 3 Calibra tions on page 88 Section 8 6 1 Cleaning the inlet filters on page 231 Monthly or when required Clean the system flow path with 1M NaOH and rinse with DM water Note Cleaning may be necessary more or less frequent ly depending on the system usage and the nature of the samples Section 8 3 2 System cleaning on page 224 Yearly or when re quried Visually inspect the drive sleeve on the Fraction collector Replace if worn out AKTA start Operating Instructions 29 0270 57 AG Maintenance Manual Chapter 7 Disassem bly 219 8 Maintenance 8 1 Regular maintenance Other required maintenance 220 Maintenance action Clean the instrument externally Clean the Fraction collec
101. eak collection is not allowed during Fractionation or Peak fractiona tion This message appears when one tries to execute Single peak collection dur ing fractionation Warning message Turning Outlet valve is not allowed during Single peak fraction ating This message is displayed to show that during Single peak fractionation the Outlet valve shall not be able to turn Occurrence message Instruction ignored Stop single peak collection is only al lowed during single peak collection This message is displayed when the in struction Single peak collection has not been executed USB removed abruptly This message appears when the user removes the USB memory stick while read or write operation is being execut ed AKTA start Operating Instructions 29 0270 57 AG 9 Troubleshooting 9 2 Basic troubleshooting USB memory stick connection To troubleshoot possible issues encountered when connecting a USB memory stick to AKTA start follow the procedure presented in the flow chart below Connect the USB memory stick to KTA start l l No Unplug and re plug the USB memory Is USB stick recognized N stick and no read write errors Is USB stick recognized No Restart the KTA start instrument Is USB stick recognized Use a new USB memory stick Is USB stick recognized Proceed with your work Contact GE Service Engineer KTA start Operating Instructions 29 0270 57 AG 25
102. ed method to a USB memory stick connected to the computer Note Make sure that the method is stored in a GE folder Method Import to AKTA start USB import Follow the instructions below to import a method from UNICORN start to AKTA start Step Action 1 2 Create a method using the Method Editor module in UNICORN start Select File Export Export Method for KTA start to USB in order to export the created method to a USB memory stick Connect the USB memory stick to AKTA start Note Make sure that the method is stored in a GE folder From the AKTA start Home screen tap Create method USB import Select the method to import AKTA start Operating Instructions 29 0270 57 AG 207 6 Operations from the Instrument Display 6 6 Manage methods and files 6 6 3 BMP result file 6 6 3 BMP result file Introduction In order to provide the user with the option to view the result image using another type of software outside of AKTA start and UNICORN start the instrument provides the feature to export the result in BMP format This format facilitates viewing of the generated result without UNICORN start Feature of exported result e The result file in BMP format can be opened and viewed using Operating Systems OS such as Microsoft Windows operating system and Mac OS e The BMP result is saved and exported if user selects the option to Save result to USB before the start of a run e The
103. ed standards Directive 2006 42 EC Machinery Directive MD 2004 108 EC Electromagnetic Compatibility EMC Directive 2006 95 EC Low Voltage Directive LVD AKTA start Operating Instructions 29 0270 57 AG 9 1 Introduction 1 3 Regulatory information CE marking The CE marking and the corresponding EU Declaration of Conformity is valid for the in strument when it is e used asa stand alone unit or e connected to other products recommended or described in the user documentation and e used in the same state as it was delivered from GE except for alterations described in the user documentation International standards This product fulfills the requirements of the following standards Standard Description EN IEC 61010 1 UL Safety requirements for EN standard is 61010 1 CAN CSA electrical equipment for mea harmonized with EU C22 2 No 61010 1 surement control and laborato directive 2006 95 EC ry use EN IEC 61326 1 Electrical equipment for EN standard is Emission according measurement control and harmonized with EU to CISPR 11 Group 1 laboratory use EMC require directive 2004 108 EC class A ments EN ISO 12100 Safety of machinery General EN ISO standard is principles for design Risk harmonized with EU assessment and risk reduction directive 2006 42 EC Regulatory compliance of connected equipment Any equipment connected to KTA start should meet the safet
104. ein and an oppositely charged chromatography medium Gel filtration GF Gel filtration also known as size exclusion chro matography is a chromatography technique that separates molecules based on differences in the molecular size Desalting buffer exchange DS Desalting is a gel filtration technique that allows rapid group separation of high molecular weight substances from low molecular weight sub stances Small molecules like salt free labels and other impurities are efficiently separated from the high molecular weight substances of interest AKTA start Operating Instructions 29 0270 57 AG 179 6 Operations from the Instrument Display 6 4 Perform a method run 6 4 3 Templates Affinity AC or lon exchange IE Follow the instruction below to start a run based on Affinity AC or lon exchange IEX 180 Note Note Step Make sure that a USB memory stick is connected to the instrument If the instrument does not detect a USB memory stick the result will not be saved Before starting the run set the status of the Fraction collector to enabled or disabled as required Action ji Method run Quick start Templates User defined Prepare system l a ED In the Method run screen tap Templates to access the different templates Templates Affinity ac C Desalting buffer exchange DS O lon exchange IEX Gel filtration GF eo A cca e Ta
105. elow WARNING WARNING indicates a hazardous situation which if not avoided could result in death or serious injury It is important not to proceed until all stated conditions are met and clearly understood CAUTION CAUTION indicates a hazardous situation which if not avoided could result in minor or moderate injury It is important not to pro ceed until all stated conditions are met and clearly understood gt NOTICE NOTICE indicates instructions that must be followed to avoid damage to the product or other equipment Notes and tips Note A note is used to indicate information that is important for trouble free and optimal use of the product Tip A tip contains useful information that can improve or optimize your procedures KTA start Operating Instructions 29 0270 57 AG 1 Introduction 1 3 Regulatory information 1 3 Regulatory information Introduction This section describes the directives and standards that are fulfilled by AKTA start Manufacturing information The table below summarizes the required manufacturing information For further infor mation see the EU Declaration of Conformity DoC document Requirement Content Name and address of manufacturer GE Healthcare Bio Sciences AB Bj rkgatan 30 SE 751 84 Uppsala Sweden Conformity with EU Directives This product complies with the European directives listed in the table by fulfilling the corresponding harmoniz
106. ement of tubing and filters 8 8 2 Replace the tubing and connectors 8 8 2 Replace the tubing and connectors Maintenance interval Replace tubing and connectors when required for example when the tubing is clogged or bent and the flow is obstructed Note Before starting to replace tubing and connectors clean the system flow path with DM water then remove the inlet tubing from water Required items The following item are required e Tubing and connectors e Tubing cutter Instruction Follow the instructions below to replace tubing and connectors Note To replace the pump tubing Marprene tubing Part No 29 0240 12 follow the instruction presented in the AKTA start Maintenance Manual Note To replace the tubing that connects the UV monitor to the Conductivity monitor use the pre bent tubing supplied with the system Step Action 1 Unscrew the connectors and disconnect the tubing 2 If the tubing has labels remove the labels and use them with the new tubing later Discard the used tubing and connectors 240 AKTA start Operating Instructions 29 0270 57 AG 8 Maintenance 8 8 Replacement of tubing and filters 8 8 2 Replace the tubing and connectors Step Action J Cut the new tubing to the same length as the original tubing Use a tubing cutter to get a straight angle cut CAUTION Cut injuries The tubing cutter is very sharp and must be handled with care to avoid injuries Note
107. ent Display 6 4 Perform a method run 6 4 2 Quick start Step Action Run on hold UV absorbance 1 0 mAU Conductivity 25 0 mS cm 2 Flow rate 3 0 ml min Graph Pressure 0 30 MPa Tube no 6 Resume End To resume the run tap Resume Note During hold the run is temporarily interrupted with current flow rate gradient and valve positions sustained 178 AKTA start Operating Instructions 29 0270 57 AG 6 Operations from the Instrument Display 6 4 Perform a method run 6 4 3 Templates 6 4 3 Templates Introduction AKTA start provides four method templates based on the most commonly used purifica tion techniques The templates are provided with default run parameters The parameters can also be changed to suit the run conditions New methods can be created and saved from these predefined templates in the Create method option This section describes how to start a run using Templates Predefined method templates The user can create customized purification methods based on the templates available on the instrument The predefined templates available with AKTA start are described below Method Description Affinity AC Affinity Chromatography separates molecules based on the reversible interaction between the target protein and the specific ligand attached to a chromatography matrix lon exchange IEX lon Exchange Chromatography is based on the reversible interaction between a charged prot
108. f the pressure reaches above the set limit the instrument will enter the Pause state Note The USB memory stick stores the result files which can be viewed using UNICORN start Also a BMP file is generated which can be viewed from any PC For details refer to Section 6 6 3 BMP result file on page 208 Do not remove the USB memory stick until the system generates the report BMP file 4 Run parameters Sample from Pump Loop Sample volume v yam ml lt gt Equilibration i j volume Oe Wash unbound oo volume e Select the mode of sample injection in the Sample from field to be ap plied via Pump or via Loop For detailed instructions on sample applica tion refer to Section 5 7 Sample application on page 135 e Set the run parameters Sample volume the volume of sample to be loaded in to the column Equilibration volume the volume of buffer A required for for equili brating the column AKTA start Operating Instructions 29 0270 57 AG 185 6 Operations from the Instrument Display 6 4 Perform a method run 6 4 3 Templates Step Action Note Washout unbound is not applicable for GF DS methods Note Make sure to load the recommended sample volume for selected column If large GF columns are used it is recommended to pre equilibrate the column before starting the run Run parameters lt l Elution volume CV une Sl OO volume e Set the run parameters
109. fer A or Buffer B Buffer B Valve position Turn valve Turn valve Switches the valve be tween its two positions i e the A and Valve switches o Counts B inlet ports Valve switches The text box displays the number of counts the valve has switched Reset Resets the counts to zero when a new valve has been re placed Sample valve Valve position Shows which inlet connection in the valve is open Buffer Buffer or Sample Sample Sample valve Valve position Turn valve Valve switches o Counts Reset Turn valve Switches the valve be tween its two positions i e the Buffer and Sample inlet ports Valve switches The text box displays the number of counts the valve has switched Reset Resets the counts to zero when a new valve has been re placed AKTA start Operating Instructions 29 0270 57 AG 55 3 System description 3 3 Instrument Display 3 3 4 Description of Settings and service Module Function Wash valve Valve position Shows which outlet z connection in the valve is open Waste Waste or Column Column EE ETF Turn valve Switches the valve be tween its two positions i e the ii cara o ci Waste and Column outlet ports Valve switches The text box displays the number of counts the valve has switched Reset Resets the counts to zero when a new valve has been re placed Outlet valve Valve position Shows w
110. ff the instrument and keep the cold cabinet open to avoid overheat ing Place the computer in room temperature If KTA start is placed in a cold room place the computer outside the cold room and use the PC Connectivity cable delivered with the in strument to connect to the computer Keep the pump cover open when not using the system Open the peristaltic pump cover after you switch off the equipment This will enhance the life time of the pump tubing KTA start Operating Instructions 29 0270 57 AG 21 2 Safety instructions 2 1 Safety precautions Maintenance WARNING To avoid damage to person when performing maintenance on AKTA start follow the instructions below e Electrical shock hazard Do not open any covers or parts unless specified in the user documentation Except for the mainte nance and service described in the user documentation all other repairs should be done by service personnel authorized by GE e Only spare parts and accessories that are approved or supplied by GE may be used for maintaining or servicing AKTA start e Disconnect power Always switch off power to the instrument before replacing any component on the instrument or cleaning the instrument unless stated otherwise in the user documen tation e Spillage Hazard Avoid spillage of fluids on the surfaces of the instrument which have cables plugs and other wirings Be careful if there is spillage of fluids on the tray while tr
111. ge Gel filtration and Desalting techniques Run parameters like column volume flow rate equilibration and elution mode and volume are predefined in the method The user needs to enter only the sample volume For detailed description of each Quick start method refer to AKTA start System Cue Card Note If required the run parameters can be changed using the Edit run option during an ongoing run Quick start techniques The table below describes the various kinds of quick start techniques that a user can choose based on application requirements Method Chromatography Details Technique AC step 1 ml 5ml Affinity Bound proteins are eluted in a single step HiTrap Chromatography using a single elution buffer Commonly used for purification of tagged proteins for example Histidine tagged proteins DS 5 ml 53ml Desalting Proteins are eluted in a single step using HiTrap a single elution buffer IEX step 1 ml 5mI lon Exchange Bound proteins are eluted in a single step HiTrap Chromatography using a single elution buffer IEX gradient 1 lon Exchange Bound proteins are eluted using two ml 5ml HiTrap Chromatography buffers with linear increase in the concen tration of buffer B over a specified time followed by a step of 100 B GF 16 60 HiPrep Gel Filtration Proteins are eluted in a single step using a single elution buffer Note Itis recommended to use the appropriate column as indicated in the te
112. greater curacy Frac30 Parameter Number of fractions Up to 30 Vessel type KTA start Operating Instructions 29 0270 57 AG Centrifuge tubes 10 to 12 ml Falcon tubes 15 ml Eppendorf tubes 1 5 ml or 2 ml 5 ml tubes 12 x 75 mm 10 Reference information 10 1 Specifications 264 Parameter Fraction volumes 0 5 to 15 ml Flammable liquids No Delay Volume UV to Dis penser head 0 49 ml default Dimensions W x D x H 270 x 280 x 285 mm Weight 5 Kg AKTA start Operating Instructions 29 0270 57 AG 10 Reference information 10 2 Chemical resistance 10 2 Chemical resistance Flow path All chemicals and concentrations are for short term lt 1 day use only ambient temper ature lt 25 C if not other stated Long term use is defined as 1 month The flow path shall withstand the following suggested chemicals Chemical Aqueous buffers pH 2 to 12 Concentration CAS no EEC no N A Usage Separation Acetone 10 67 64 1 200 662 2 Acetic acid 6 1 M 64 19 7 200 580 7 CIP Ammonium sulphate 35M 77 83 20 2 231 984 1 Purification of plasmids Arginine 2 M 74 79 3 200 811 1 Wash using pro tein A gels Refold ing Benzyl alcohol 100 51 6 202 859 9 Cleaning and storage of columns Decon 90 1310 58 3 215 181 3 Cleaning Dimethyl sulphoxide DMSO 67 68 5
113. he Injection valve is in Load position before starting the run 6 After manually switching position acknowledge the message by tapping Continue The sample will be injected onto the column when the Injection valve is switched manually to position Inject to column during the run 7 Switch the Injection valve position to the Load position when the following screen is shown on the Instrument Display Sample inject Turn the Manual Injection Valve to Load position and then tap Continue 8 After manually switching the position of the Injection valve acknowledge the message by tapping Continue Note For Binding techniques AC IEX it is advisable to empty the loop with at least 3 times the loop volume to achieve good high sample recovery This is not advised for non binding techniques DS GF as there are sample volume limi tations due to the size of the column used AKTA start Operating Instructions 29 0270 57 AG 143 5 Prepare the system for a run 5 7 Sample application Load the sample from a Superloop Superloop allows the injection of larger volumes of sample 10 to 150 ml onto the column Follow the instructions below to apply the sample using a Superloop Step Action 1 Fill a large volume syringe with sample 2 Connect the syringe to the Injection valve port 3 and carefully inject the Sample into the Superloop Note Make sure that the Injection valve is set to position Load sample which al lows to fi
114. hich outlet connection in the valve is open Waste Waste or Collection Collec tion Outlet valve Valve position Turn valve Turn valve Switches the valve be tween its two positions i e the Valve switches Counts i E Waste and Collection Frac outlet ports A Valve switches The text box displays the number of counts the valve has switched Reset Resets the counts to zero when a new valve has been re placed 56 KTA start Operating Instructions 29 0270 57 AG Settings and service Screen 3 The options available for the modules displayed on Settings and service Screen 3 are briefly described below Module Main board Main board Main board Power supply Memory test Temperature FPGA test Back n V C 3 System description 3 3 Instrument Display 3 3 4 Description of Settings and service Function Power supply Shows the actual voltage Memory test Performs a memory test Temperature Shows the tempera ture of the main board FPGA test Performs a test of the FPGA hardware UV UV UV LED calibration Diagnostics Back _ Flow cell path length Configuration UV LED calibration To calibrate theUV LED intensity Flow cell path length To derive the actual path length of the flow cell Diagnostics To troubleshoot the UV module Configuration
115. hod 5 4 1 System performance method Introduction The System performance method is performed to make sure that the system is perform ing within acceptable limits It is recommended to run the test at the time of installation of the instrument or after replacement of modules such as Pump UV Conductivity or Valves System performance method can also be used at any time to check the condition of the system for example after a prolonged storage of the system The System perfor mance method can be performed from both the Instrument Display and UNICORN start Note e Calibrate all the modules before starting the test e Make sure that there is no column connected e tis recommended not to edit any run parameters during a test in order to avoid the failure of the test Requirements Required solutions are e Buffer A DM water e Buffer B 1 0 acetone 1 0 M NaCl e Sample 1 0 acetone 1 0 M NaCl Buffer B Note Make sure to prepare the buffer solutions accurately to avoid the test failure Checklist Before starting a System performance method make sure that the following tasks are completed or met e Calibration of all the modules Pressure sensor Pump UV and Conductivity e Nocolumn should be present in the flow path e Set the conductivity reference temperature to 20 C save and enable the function e Immerse Buffer port A inlet in Buffer A DM water e Immerse Buffer port B inlet in Buffer B 1 0 acetone 1 0
116. ich can be viewed from any PC For details refer to Section 6 6 3 BMP result file on page 208 Do not remove the USB memory stick until the system generates the report bmp file Note USB imported methods which were created in UNICORN start cannot be edited in the instrument Use UNICORN start to edit those methods AKTA start Operating Instructions 29 0270 57 AG 6 Operations from the Instrument Display 6 4 Perform a method run 6 4 5 Prepare system methods 6 4 5 Prepare system methods Introduction Predefined methods for the preparation and the cleaning of the system are available with AKTA start Use the Prepare system methods to clean the entire system flow path when needed and to fill the system with storage solution when the instrument is not going to be used for a period of time For detailed instructions see Section 8 3 Cleaning the system flow path on page 222 The system methods available with AKTA start are listed below e Pump washA e Pump wash B e Washout fractionation tubing e Column preparation e System cleaning e System performance method The Pump wash A B Washout fractionation tubing and Column preparation methods required for the system preparation are presented in detail in Section 5 6 Run Prepare system methods on page 125 The System performance method is presented in Section 5 4 System performance on page 106 System cleaning Follow the instruction below to perform a system cleaning
117. in flow cell to avoid damage Rinse the flow path with water thoroughly e Make sure that the NaOH is completely removed The conductivity reading in the Run view screen should be lt 1 mS cm AKTA start Operating Instructions 29 0270 57 AG 229 8 Maintenance 8 6 Other cleaning procedures 8 6 Other cleaning procedures Introduction This section provides instructions for additional cleaning procedures to be performed by the user of AKTA start In this section This section contains the following subsections Section See page 8 6 1 Cleaning the inlet filters 231 8 6 2 Cleaning the instrument externally 232 8 6 3 Cleaning the Fraction collector 233 KTA start Operating Instructions 29 0270 57 AG 8 Maintenance 8 6 Other cleaning procedures 8 6 1 Cleaning the inlet filters 8 6 1 Cleaning the inlet filters Maintenance interval Clean the inlet filters when required for example when the visual inspection shows that the filters are clogged Required solutions The following solutions are required e 1M NaOH e DM water Instruction Follow the instructions below to clean the inlet filters Step Action 1 Pull off the support net and the inlet filter from the inlet filter holder See Section 8 8 1 Replace the inlet filters on page 239 2 Immerse and leave the inlet filter and the support net in 1 M NaOH for about 2 hours 3 Remove the inlet filter and the support net from the NaOH solutio
118. ing buffer exchange DS C lon exchange IEX Gel filtration GF a Lo e Tapa radio button to select a template e Tap Create to create a method based on the selected technique For details on the templates available in AKTA start refer to Section 6 4 3 Templates on page 179 Set parameters Column volume v ym mi Flow rate v ym ml min r Pressure limit v ym MPa Save method as RGO 1 3 9 e Set the run parameters Column volume column volume ml Flow rate flow rate ml min Pressure limit pressure limit MPa AKTA start Operating Instructions 29 0270 57 AG 197 6 Operations from the Instrument Display 6 6 Manage methods and files 6 6 1 Create method Step Action e Select Save Method as field if you want to set a method name The file name can be edited by setting the digits in the range 00 to 99 Note Provide a unique method name The created method will be saved under the User defined methods menu Note Make sure that the values for the Column volume Flow rate and Pressure limit are appropriate for the chosen column Refer to the column manual for details If the pressure reaches above the set limit the instrument will enter the Pause state 4 Set parameters 2 Sample from Pump Loop Sample volume ml Equilibration CV Wash unbound CV 2 3 a E e Select the mode of sample injection in the Sample from field to be ap plied via Pump or
119. ing if required Is the buffer inlet tubing immersed in correct buffer solutions See Section 5 10 Starting a run on page 153 Are the inlet filters clean Clean the inlet filters or replace the filters if required See Section 8 6 1 Cleaning the inlet filters on page 231 and Section 8 8 1 Replace the inlet filters on page 239 Is the column connected correctly See Section 5 5 Connect a column on page 121 Purification checks 244 Has the column been cleaned and prepared according to the recommendations in the column instruction manual Has the sample been adjusted to binding buffer conditions Has the sample been clarified by centrifugation and or filtration prior to the sample loading Are the correct buffers used for the chosen columns and proteins Check buffers for precipitations and contamination Adjust the required temperature wherever needed Are the chosen columns suitable for the chosen target proteins Have the stable baseline for UV and Conductivity been established AKTA start Operating Instructions 29 0270 57 AG 9 Troubleshooting 9 2 Basic troubleshooting 9 2 Basic troubleshooting Introduction This section describes problems that can occur with the Display UV and Conductivity measurements problems with the wet side modules possible causes and the recom mended corrective actions If error codes are displayed on the Touch screen refer to AKTA start Maintenance Manual Display Des
120. ion This section describes the types of methods that can be selected for a run In this section This section contains the following subsections Section See page 6 4 1 Select a method type 171 6 4 2 Quick start 173 6 4 3 Templates 179 6 4 4 User defined methods 187 6 4 5 Prepare system methods 189 170 AKTA start Operating Instructions 29 0270 57 AG 6 4 1 Select a method type Method types 6 Operations from the Instrument Display 6 4 Perform a method run 6 4 1 Select a method type Four different method types can be selected to perform a method run The different method types are specified below Quick start Allows the user to run methods like affinity lon exchange Gel filtration and Desalting with method parameters predefined Templates Allows the user to edit and run the predefined methods Affinity lon exchange Gel filtration and Desalting User defined Allows the user to run user created methods or USB imported methods Prepare system Allows the user to perform system operations such as Pump wash Column preparation Cleaning and System performance test The Quick start methods and Templates available with AKTA start are briefly described in Section 6 4 2 Quick start on page 173 and Section 6 4 3 Templates on page 179 For a description of the Prepare system methods see Section 6 4 5 Prepare system methods on page 189 and Section 8 3 Cleaning the system flow path on page 222 Method type Q
121. ion Weigh beaker 4 2 to 5 89 End gradient stay at 100 B Gradient 5 Straight no nega tive dips End Check all connec No leakage tions for leakage 1 With Fraction collector 2 Without Fraction collector 3 UV 50 B UV 100 B Note e Make sure to update the Performance result text file with manually ob served recordings and then print the report e The other parameters are automatically checked and pass fail reports are generated in the report For detailed list of acceptance criteria refer to Section 5 4 2 System performance method from AKTA start on page 109 AKTA start Operating Instructions 29 0270 57 AG 115 5 Prepare the system for a run 5 4 System performance 5 4 3 System performance method from UNICORN start Illustration of the System performance test The illustration below represents a typical System performance method result file gen erated from UNICORN start 280 Note 4 UV 4 Conductivity 4 Gradient Concentration System Flow Pressure Fraction Run Log mi 10 15 20 25 30 35 40 45 50 55 It is recommended to optimize the timing of switch valve when wavy gradients or fluctuations are observed For detailed description see Section 5 4 4 Switch valve timing on page 117 If the System performance method fails analyze the cause for the failure based on the acceptance criteria Perform the following actions e Re calibrate the failed modu
122. ion collector to enabled or disabled as required AKTA start Operating Instructions 29 0270 57 AG 183 6 Operations from the Instrument Display 6 4 Perform a method run 6 4 3 Templates Step Action 1 Method run In the Method run screen tap Templates to access the predefined method templates 2 Templates O Affinity AC O Desalting buffer exchange DS lon exchange IEX Gel filtration GF Back ES EZS e Tapa radio button to select a template that suits your application e g Gel filtration e To continue with the selected technique tap Select 3 Run parameters Column volume v 1200 Flow rate v i ym ml min D 030 7 Pressure limit v 4 to USB He 184 AKTA start Operating Instructions 29 0270 57 AG 6 Operations from the Instrument Display 6 4 Perform a method run 6 4 3 Templates Step Action e Set the run parameters Column volume column volume ml Flow rate flow rate ml min Pressure limit pressure limit MPa Use up down arrows to set the values or use numpad to type in the values e Tick the checkbox Save Result to USB if you want to save the result The result file name can be edited by setting the digits in the range 00 to 99 e Tap the forward arrow to access additional run parameters Note Make sure that the values for the Flow rate and Pressure limit are appropriate for the chosen column Refer to the column manual for details I
123. ion collector using Outlet valve and in between different runs using the Fraction collector Note Flow is diverted from fractionation tubing to collection tube through Outlet valve Requirements Required cleaning solutions are e DM water e Buffer solution Follow the instructions below to perform a Washout fractionation tubing run The Washout fractionation tubing procedure is initiated from the Instrument Display Instruction Step 1 2 3 4 130 Action Immerse the buffer inlet tubing in DM water or buffer Remove the column from the flow path and re connect the flow path For detailed instructions see Section 8 3 1 Disconnect the column on page 223 Place the fractionation tubing in the waste container In the KTA start home screen tap Method run KTA start 2 Create method Settings and service AKTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 6 Run Prepare system methods 5 6 3 Washout fractionation tubing Step Action 5 In the Method run screen tap Prepare system Method run 2 Quick start Templates User defined Prepare system A 6 Select Washout fractionation tubing and then tap Select to initiate the method Select system method 2 D Pump wash A Pump wash B gt Washout fractionation tubing O Column preparation 1 2 co o EB 7 Set the required run parameters e Flow rate flow rate ml min e
124. ispens assembly ing the liquid into fractions Tubing holder Holds the tubing used for dispensing the liquid fractions into the collection tubes Collection tubes 10 to 18 mm diameter tubes used to collect the fractions Bowl assembly Holder for collection tubes which supports tubes of four sizes Base unit Case for electromechanical assembly and holder for the Bowl assembly Power on indicator 36 AKTA start Operating Instructions 29 0270 57 AG 3 System description 3 1 System overview Main features of AKTA start The main features of AKTA start are listed below KTA start is a compact and one step purification solution for quick and reliable pu rification of proteins A simple and modern system offered to automate the protein purification workflow by providing features like automated sample injection fraction collection real time monitoring Method templates are available for all common chromatography techniques such as Affinity Chromatography lon Exchange Chromatography Gel filtration and De salting Quick start methods are available for purifying several common proteins Predefined system methods are available for cleaning the flow path AKTA start is operated using a touch screen on the instrument In addition the system can be operated from a computer connected to the instrument using the UNICORN start software KTA start is offered with a dedicated Fraction collector Frac30 al
125. keep a minimum number of files on the memory stick Once you take a backup of the files delete them from the memory stick and then save onto a computer Avoid keeping unnecessary files in the memory stick Avoid using folders named GE on the USB memory stick However you may use folders named for example GE_ or GExyz Always take backup of complete GE folder from USB memory stick and do not backup individual files It is recommended to take backups whenever you have completed important runs Result file import from USB stick to UNICORN start Follow the instructions below to export a result file generated in AKTA start and import it into UNICORN start Step Action 1 2 Open the Evaluation module in UNICORN start Select File Import Import KTA start results from USB and then import the result files to a desired location on the computer View analyze report or print the result file Method Export from UNICORN start to USB memory stick Follow the instructions below to export a method created in UNICORN start to a USB memory stick Step Action 1 2 Create a method using the Method Editor module in UNICORN start Connect a USB memory stick to the computer AKTA start Operating Instructions 29 0270 57 AG Step 6 Operations from the Instrument Display 6 6 Manage methods and files 6 6 2 Handling the USB memory stick Action Select File Export Export Method in order to export the creat
126. lay e Conductivity Procedures required after a run such as the cleaning of the column the system flow path can also be performed manually or using methods available in the Prepare system menu 156 AKTA start Operating Instructions 29 0270 57 AG 6 Operations from the Instrument Display 6 1 Introduction Calibration of the modules and service can be performed from the Settings and service screen Detailed instructions for calibration are presented in Section 5 3 Calibrations on page 88 Method management operations such as create edit and import a method can be performed from the Create method screen For detailed instructions see Section 6 6 Manage methods and files on page 195 Operation Overview The AKTA start home screen displays four different options for the user to select and perform operations from Instructions for each operation are presented in separate sections in this chapter For a description of the options available in the main screen see Chapter 3 System description on page 33 AKTA start Create method Settings and service Checklist Make sure that the system is correctly prepared Check that e The system is prepared according to Chapter 5 Prepare the system for a run on page 83 and the modules are calibrated according to Section 5 3 Calibrations on page 88 e A suitable column has been selected for the application Consider target protein pressure range and optimal flow
127. le Step Action 1 Fill a syringe with sample AKTA start Operating Instructions 29 0270 57 AG 141 5 Prepare the system for a run 5 7 Sample application Step Action 2 Connect the syringe to the Injection valve port 3 INJEGI ION VALVE Note Make sure that the Injection valve is set to position Load Sample which al lows to fill the sample loop from the fill port 3 3 Carefully load the sample into the sample loop To avoid sample loss due to siphoning leave the syringe in the port until the sample has been injected onto the column during the run Tip It is recommended to overfill the loop to make sure that the loop is completely filled Excess of sample will leave the valve through port 4 4 From the Instrument Display select Method run then choose Templates For details see Section 6 4 Perform a method run on page 170 142 AKTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 7 Sample application Step Action 5 In the Run parameters screen select the sample application via the Loop and set all the required run parameters For details see Chapter 6 Operations from the Instrument Display on page 155 Switch the Injection valve position to the Inject position when the following screen is shown on the Instrument Display Sample inject Turn the Manual Injection Valve to Inject position to inject the sample and then tap Continue Note Make sure that t
128. le e Use buffer with proper composition e Clean the failed module or entire system Refer Chapter 8 Maintenance on page 217 for more details on cleaning e Carefully follow the test instructions e Repeat the System performance method until it passes e fwavy or fluctuations in gradient is observed then perform Switch valve timing optimization e Ifthe test fails after following the above actions replace the failed module 116 AKTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 4 System performance 5 4 4 Switch valve timing 5 4 4 Switch valve timing Introduction Switch valve timing is used to optimize the switch valve Buffer valve timing of AKTA start It is recommended to optimize the timing of switch valve when wavy gradients are obtained or when fluctuations in the step gradient are observed during either System performance method or chromatography runs Note The default Switch valve timing A is 4 sec Switch valve timing B is 5 sec Switch valve time can be set between 3 0 and 5 0 sec with 0 1 sec incre ments using Advanced timing The gradient fluctuations or wavy gradients are highly dependent on the flow rates It is recommended to modify the switch valve timing if wavi ness fluctuations are seen for specific flow rates After changing the Switch valve time perform a System performance method to evaluate the gradient fluctuations wavy gradients Alternatively a man
129. leaning 8 3 2 System cleaning Introduction The System cleaning method is used to clean the instrument flow path System cleaning is recommended to be performed to prevent carryover between runs contamination in the flow path as a routine maintenance protocol and to prepare system for storage Note e Cleaning is important for preventing cross contamination and bacterial growth in the instrument e Prepare cleaning solutions of recommended concentration to ensure proper cleaning e tis recommended not to end the run before completion e tis recommended to clean the inlets and outlets sample tubing fraction ation tubing from the Edit run screen Required solutions Instruction 224 The following cleaning solutions are required e 1MNaOH e DM water Follow the instructions below to clean the system flow path The System cleaning pro cedure is initiated from the Instrument Display Step Action 1 Remove the column from the flow path and re connect the tubing 2 Immerse both the buffer inlets in 1M NaOH 3 In the KTA start home screen tap Method run KTA start 2 Create method Settings and service AKTA start Operating Instructions 29 0270 57 AG Step 8 Maintenance 8 3 Cleaning the system flow path 8 3 2 System cleaning Action In the Method run screen tap Prepare system Method run Quick start Templates User defined Prepare system EI Select System
130. leaning the UV flow cell Step Action 2 Start a manual run and pump cleaning solution at 5 ml min through the UV flow cell for 10 minutes and pause the run 3 Leave the UV flow cell filled with cleaning solution for 15 minutes 4 Immerse the inlet tubing in DM water 5 Resume the run and rinse the flow cell thoroughly with DM water 228 AKTA start Operating Instructions 29 0270 57 AG 8 Maintenance 8 5 Cleaning the Conductivity flow cell 8 5 Cleaning the Conductivity flow cell Maintenance interval Clean the Conductivity flow cell when the Conductivity Monitor shows a slow response or when the conductivity measurements are not comparable to any of the previous re sults Required solutions The following solutions are required e 1MNaOH e DM water Cleaning the Conductivity flow cell in place Follow the instructions below to clean the Conductivity flow cell Note Before cleaning the Conductivity flow cell remove the column from the flow path and re connect the flow path See Section 8 3 1 Disconnect the column on page 223 Step Action 1 Immerse the inlet tubing either A or B in 1M NaOH Start a manual run and pump 1M NaOH at 1 ml min through the flow cell for 10 minutes 3J Pause the run Leave the Conductivity flow cell filled with 1M NaOH for 15 minutes 4 Immerse the inlet tubing in DM water 5 Resume the run and rinse the flow cell thoroughly Note e Donotleave NaOH for avery long time
131. level 7 1 ml min Sample valve Buffer position 10 Request switch Injection Max UV level 300 to 380 MAU valve to Inject position Max Conductivi 65 to 95 mS cm ty level 13 Request switch Injection valve back to Load posi tion 15 Start gradient 0 t0100 B in 10 minutes start fraction ation collection 19 End fractionation 1 Weigh fraction 0 8 to 1 2 g no 2 3 and 4 Max diff be 0 1g tween fractions 276 KTA start Operating Instructions 29 0270 57 AG Activity End collection Weigh beaker 11 Appendix 11 1 System Performance Report Approved interval 4 2 to 5 89 Observations End gradient stay at 100 B Gradient Straight no negative dips 50 B Gradient level gt 45 to 55 B 0 B Re equilibration End 1 With Fraction collector 2 Without Fraction collector 3 UV 50 B UV 100 B AKTA start Operating Instructions 29 0270 57 AG Check all connec tions for leakage No leakages 211 11 Appendix 11 1 System Performance Report Test performed from UNICORN start Activity 1 ml min 0 B flow through Outlet valve waste position Repeat UV Auto zero Back pressure Approved inter val Observations 5 ml min Back pressure 0 06 to 0 2 MPa Start gradient O to 100 B in 10 minutes start fraction ation collection End fractionation 1 Weigh fraction no 2 3 and 4 0 8 to 1 2g Max difference betwee
132. librated if equilibration is not included in the method Sample e Make sure that the sample is ready to be loaded via Pump Loop or Superloop Pump e Make sure to place the pump tubing properly over the pump head Make sure to close the pump hood before starting the run Result storage e f the result of the run has to be saved when using without UNICORN start make sure that a USB memory stick is connected to the instrument UNICORN start e Check that KTA start is connected to a PC with UNICORN start installed e Make sure that the system connection Is established before starting the run For details refer to UNICORN start 1 0 User Manual AKTA start Operating Instructions 29 0270 57 AG 153 5 Prepare the system for a run 5 10 Starting a run Start a run 154 A chromatography run is performed on KTA start is by either using a Quick start method or Template or by running the system manually A run can be started from the Instrument Display or from UNICORN start by selecting one of the run options available with the in strument Detailed instructions for starting a run are presented in Chapter 6 Operations from the Instrument Display on page 155 For starting a run from UNICORN start refer to UNICORN start 1 0 User Manual AKTA start Operating Instructions 29 0270 57 AG 6 Operations from the Instrument Display 6 Operations from the Instrument Display About this chapter This chapter describes ho
133. ll the capillary loop from the fill port 3 x From the Instrument Display select Method run then choose Quick start methods or Templates For details see Section 6 4 Perform a method run on page 170 4 In the Run parameters screen select the sample application via the Loop and set all the required run parameters For details see Operation Overview on page 157 144 AKTA start Operating Instructions 29 0270 57 AG Step 5 Prepare the system for a run 5 7 Sample application Action J Switch the Injection valve position to the Inject position when the following screen is shown on the Instrument Display Sample inject Turn the Manual Injection Valve to Inject position to inject the sample and then tap Continue Note Make sure that the Injection valve is in Load position before starting the run After manually switching position acknowledge the message by tapping Continue Switch the Injection valve position to the Load position when the following screen is shown on the Instrument Display Sample inject lN Turn the Manual Injection Valve to Load position and then tap Continue The sample will be injected onto the column when the Injection valve is switched manually to the position Inject to column during the run KTA start Operating Instructions 29 0270 57 AG 145 5 Prepare the system for a run 5 7 Sample application Step Action 8 After manually switching the position of the
134. lowing to collect the fractions in four different tube sizes AKTA start Operating Instructions 29 0270 57 AG 37 3 System description 3 2 Instrument 5 2 Instrument Introduction This section provides an overview of AKTA start Illustration of the instrument modules The illustration below shows the locations and gives brief descriptions of the modules placed on the wet side of the instrument Part Function 1 Buffer valve 9 10 Description A 3 port valve that is used as a switching valve for gradient formation It enables the use of two buffers which are required for forming a gradient during runs 38 AKTA start Operating Instructions 29 0270 57 AG Part 2 Function Mixer Sample valve 3 System description 3 2 Instrument Description A static mixer that is used for mixing of buffers A and B A 3 port valve that allows either the buffer or the sample to enter the flow path The Sample valve enables direct application of the sample onto the column using the Pump Pump A peristaltic pump which delivers buffer or sample to the flow path with a flow rate of up to 5 ml min For cleaning procedures the Pump can flush the flow path at a flow rate of 10 ml min Pressure sensor The Pressure sensor reads the pressure in the flow path and senses overpressure Wash valve A 3 port valve that is used to divert the flow path to waste The Wash valve switches automaticall
135. m configuration Benchtop system Control system Instrument Display and or UNICORN start Connection between PC and instrument USB Dimensions W x D x H 340 mm x 280 mm x 360 mm Weight excluding packaging 8 kg Power supply 100 to 240 V AC 10 50 60 Hz Power consumption 95 VA Transient overvoltages Overvoltage category Fuse Fast blow glass tube type FSAL250V Enclosure protective class IP21 Tubing and connectors Inlet Buffer valve to Mixer PTFE tubing length 100 cm i d 1 6 mm 5 16 24 UNF connections PEEK tubing Length 15 cm i d 0 75 mm 10 32 UNF connections Mixer to Sample valve PEEK tubing Length 23 cm i d 0 75 mm 10 32 UNF connections Sample valve to Pressure sensor via Pump Marprene tubing Length 25 cm i d 0 8 mm 10 32 UNF connections Pressure sensor to Wash valve PEEK tubing Length 13 cm i d 0 75 mm 10 32 UNF connections KTA start Operating Instructions 29 0270 57 AG 10 Reference information 10 1 Specifications Parameter Wash valve to Injection valve PEEK tubing Length 17 cm i d 0 75 mm 10 32 UNF connections Injection valve to Column PEEK tubing Length 15 cm i d 0 75 mm 10 32 UNF connections Column to UV PEEK tubing Length 15 cm i d 0 75 mm 10 32 UNF connections UV to Conductivity PEEK tubing Length 20 cm i d 0 75 mm 10 32 UNF connections Cond
136. m operation Maintenance General precautions WARNING Always follow these General precautions to avoid injury when using AKTA start e Do not operate KTA start in any other way than described in the AKTA start user documentation e Operation and user maintenance of KTA start should be per formed according to the instructions described in AKTA start Operating Instructions and AKTA start Maintenance Manual e Do not use any accessories not supplied or recommended by GE e Do not use KTA start if it is not working properly or if it has suffered any damage for example damage to the power cord its plug or the Frac30 cable damage caused by dropping the equipment damage caused by splashing liquid onto it 16 AKTA start Operating Instructions 29 0270 57 AG 2 Safety instructions 2 1 Safety precautions Using flammable liquids WARNING When using flammable liquids with AKTA start follow these precau tions to avoid any risk of fire or explosion e Fire Hazard Before starting the system make sure that there is no unintentional leakage in the instrument or tubing e Explosion hazard To avoid building up an explosive atmo sphere when using flammable liquids make sure that the room ventilation meets the local requirements CAUTION To avoid hazardous situations when unpacking installing or moving AKTA start follow the precautions below KTA start is filled with den
137. m peak integration analysis e Create and print PDF reports e Import results from KTA start via a USB memory stick AKTA start Operating Instructions 29 0270 57 AG 215 7 Operations from UNICORN start 7 4 Administration 7 4 Administration Introduction The Administration module is used to manage the UNICORN start database and to review UNICORN start and system logs Illustration of the Administration module 2 Administration la El amp File Reports Tools Help ld UNICORN and System Log Ee Database Management Main features The main features of the Administration module are listed below e Ability to back up and restore operations and archive and retrieve operations e Review the UNICORN start and system logs 216 AKTA start Operating Instructions 29 0270 57 AG 8 Maintenance 8 Maintenance About this chapter This chapter describes the maintenance program for AKTA start and provides instructions for routine maintenance In this chapter This chapter contains the following sections Section See page 8 1 Regular maintenance 218 8 2 Cleaning before planned maintenance service 221 8 3 Cleaning the system flow path 222 8 4 Cleaning the UV flow cell 227 8 5 Cleaning the Conductivity flow cell 229 8 6 Other cleaning procedures 230 8 7 Storage of the instrument 236 8 8 Replacement of tubing and filters 238 AKTA start Operating Instructions 29 0270 57 AG 217 8 Maintenance
138. m temperature AKTA start Operating Instructions 29 0270 57 AG 151 5 Prepare the system for a run 5 9 Cold room operations Step Action 3 Allow the instrument to stabilize at room temperature for at least a few hours 4 Tighten all connections and pump DM water through the system to check for leaks 5 Tighten any leaking connector 152 KTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 10 Starting a run 5 10 Starting a run Final checks Before starting a run make the checks recommended below to prevent that problems are not encountered once the run has been started Buffer e Check that the buffer inlet tubings A and B are immersed in the correct bottles con taining the buffers of interest e Check that there is sufficient buffer available Waste outlet e Check that the outlet tubings leading to waste from the Wash valve Injection valve and the Outlet valve are placed in the waste container e Check that the waste container is not full and that there is provision for the volume diverted to it during the run Fraction collector e f the fraction collector Frac30 is going to be used during the run check that the fraction collector is prepared and filled with collection tubes and the Outlet valve collection PEEK tubing is connected to the Fraction collector and also that the Fraction collector is enabled Column e Check that the correct column has been connected and equi
139. method Ability to generate a System error report Note When the Fraction collector is enabled the process picture will display a Fraction collector image If the Fraction collector is disabled a collection beaker image is displayed 212 AKTA start Operating Instructions 29 0270 57 AG 7 Operations from UNICORN start 7 2 Method Editor 7 2 Method Editor Introduction The Method Editor module provides flexibility to create or edit the chromatography methods Illustration of the Method Editor module The user interface of Method Editor is illustrated below bembea ae il E E jo E oE Pegs pel L Pii iee a o T F Er Wg ne Ba a oe z e EEN BLN ia arini Em E fee i mem n i i urme ae do ra Perr mai 2 gue ri f i i i E T i iy a Main features The main features of the Method Editor module are listed below e Ability to create methods from predefined templates like Affinity lon Exchange Desalting and Gel Filtration e Flexibility to create customized methods by dragging and dropping chromatography phases such as Prime and Equilibration Sample Application Wash Out Unbound Elution and Fractionation AKTA start Operating Instructions 29 0270 57 AG 213 7 Operations from UNICORN start 7 2 Method Editor e The methods created from Method Editor can either be run directly from System Control or exported to a USB memory stick to run them from AKTA start directly
140. mplate names For example use HiTrap 1 ml column when selecting AC IEX step1 ml HiTrap or 5 ml column when selecting AC IEX step 5ml HiTrap KTA start Operating Instructions 29 0270 57 AG 173 6 Operations from the Instrument Display 6 4 Perform a method run 6 4 2 Quick start Start a run Follow the instruction below to start a run based on a Quick start method Note Make sure that a USB memory stick is connected to the instrument If the instrument does not detect a USB memory stick the result will not be saved Step Action 1 Method run 2 In the Method run screen tap Quick start to access the templates 2 Quick start AC step 1 ml HiTrap C AC step 5 ml HiTrap gt gt O DS 5 ml HiTrap D C DS 53 ml HiPrep 1 3 Back A EZS Quick start O IEX step 1 ml HiTrap C IEX step 5 ml HiTrap lt gt D IEX gradient 1 ml HiTrap C IEX gradient 5 ml HiTrap 2 3 Back or EZS 174 KTA start Operating Instructions 29 0270 57 AG 6 Operations from the Instrument Display 6 4 Perform a method run 6 4 2 Quick start Step Action Quick start GF 16 60 HiPrep 3 3 Back Q Select e To selecta Quick start method tap a radio button or Tap the forward arrow to access additional Quick start methods e To continue with the selected method tap Select Note Make sure to load the recommended sample volume for the selected column If la
141. n and rinse thoroughly with DM water 4 Fit the inlet filter into the support net and press it into position on the inlet filter holder AKTA start Operating Instructions 29 0270 57 AG 231 8 Maintenance 8 6 Other cleaning procedures 8 6 2 Cleaning the instrument externally 8 6 2 Cleaning the instrument externally Maintenance interval Clean the instrument externally when required Do not allow spilled liquid to dry on the instrument Required material The following materials are required e Cleaning cloth e Mild cleaning agent or 20 ethanol Instruction Follow the instructions below to clean the instrument externally Step Action 1 Check that no run is in progress 2 Switch off the instrument 3 Wipe the surface with a damp cloth Wipe off stains using a mild cleaning agent or 20 aqueous ethanol 4 Let the instrument dry completely before using it 232 KTA start Operating Instructions 29 0270 57 AG 8 Maintenance 8 6 Other cleaning procedures 8 6 3 Cleaning the Fraction collector 8 6 3 Cleaning the Fraction collector Maintenance interval Clean the fraction collector when required for example in case of liquid spill in the Bowl assembly Required material The following materials are required for cleaning the Bowl assembly e Water e 20 ethanol e Cleaning cloth Instruction Follow the instructions below to disassemble and clean the Bowl assembly Step Action
142. n fractions 0 1g End collection Weigh beaker 4 2 to 5 89 End gradient stay at 100 B Gradient gt Straight no nega tive dips End 1 With Fraction collector 2 Without Fraction collector 3 UV 50 B UV 100 B 278 Check all connec tions for leakage No leakage AKTA start Operating Instructions 29 0270 57 AG Index A AKTA start instrument main parts 35 instrument modules descrip tions 38 intended use 34 main features 37 system description 34 B BMP result file 208 C Calibrations calibration guide 89 CE conformity 9 CE marking 10 Chromatogram view 165 Cleaning Conductivity flow cell 229 Inlet filters 231 UV flow cell 227 Clean the system 192 Cold room operations 151 Column connect 122 disconnect 223 placement 121 Column preparation 133 Conductivity flow cell calibration 103 cleaning 229 Conductivity Monitor reference temperature 101 set cell constant 100 temperature sensor calibra tion 102 Create method 196 E Emergency procedures emergency shutdown 28 AKTA start Operating Instructions 29 0270 57 AG Index power failure 29 Evalutate a run 192 Export method 206 F Flow path inlet and outlet tubing 85 Frac30 delay volume setting 149 main parts 36 preparation 147 Fractionation 159 Fraction collector cleaning 233 Import method 207 result file 206 Injection valve description 136 Installati
143. nected to the instrument For details about exporting a method see Sec tion 6 6 2 Handling the USB memory stick on page 205 Step Action I Create method rae ee t In the Create method screen tap USB Import to access the methods 202 AKTA start Operating Instructions 29 0270 57 AG 6 Operations from the Instrument Display 6 6 Manage methods and files 6 6 1 Create method Step Action Select method to import from USB Method 1 Method 2 a a impor e Tapa radio button to select a method e Tap Import to import the method Result A Message screen that requires to confirm the action will be dis played Message A Are you sure you want to import the method Yes No Tap Yes to confirm the import of the selected file or Tap No to cancel the action and return to the file list Note The imported methods will be saved under the User defined methods menu Only one method can be imported at a time If multiple methods are to be imported repeat the steps described above Note If the system memory is full delete existing methods before importing a new method AKTA start Operating Instructions 29 0270 57 AG 203 6 Operations from the Instrument Display 6 6 Manage methods and files 6 6 1 Create method Delete a method 204 Follow the instruction below to delete a user method Step Action 1 Create method L Create Edit EJ In
144. ngth screen e set the Cell path length in the Cell field corresponding to the new UV flow cell Note The Cell path length is provided on the packing of the UV flow cell e tap Save Path Length to save the value 4 If the Cell path length is not available follow the instructions below to cal culate the Cell path length Immerse the inlet buffer A tubing in DM water and the inlet buffer B tubing in 1 acetone solution 5 In the Flow Cell path length screen set the Cell value to 2 00 mm by using the up down arrows or typing in the value 6 Flush thoroughly and leave the UV flow cell filled with buffer A DM water Tap Set Baseline to capture a new reference for the test Result Abs should show a value close to 0 MAU 7 Flush the UV flow cell with Buffer B 1 acetone and leave the flow cell filled with 1 acetone Note down the new Abs value 8 Calculate the actual Cell path length Cell rm 2 00 new Abs value 340 AKTA start Operating Instructions 29 0270 57 AG 97 5 Prepare the system for a run 5 3 Calibrations 5 3 5 UV Monitor calibration Step Action 9 Update the calculated Cell length value by using up down buttons or typing in the value Tap Save Path Length to save the value to permanent memory Note Observe that the Abs value now should be about 340 5 mAU confirming that normalization has been done Calibrate the UV LED Follow the instructions below to calibrate the UV LED
145. no trapped air bubbles in the tubing e Prefill the sample tubing with sample before the start of the run to ensure that the tubing is filled with sample In the Run parameters screen set the sample volume and the other required parameters For details see Operation Overview on page 157 Tap Run to start the run Prime the sample loop before injecting sample A sample loop allows the injection of small sample volumes onto the column The sample application via the loop is performed in two steps 1 Loading the sample loop with sample 2 Injecting the sample from the sample loop onto the column Follow the instructions below to prime the sample loop using DM water Buffer before injecting the sample using the manual Injection valve Step Action 1 140 Fill a syringe with DM water buffer Note Make sure that the Injection valve is set to position Load Sample KTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 7 Sample application Step Action 2 Connect the syringe to the Injection valve port 3 3 Load the DM water buffer into the sample loop 4 Repeat steps 1 to 3 using at least 5 times the loop volume before loading the sample Load the sample into the sample loop Follow the instructions below to load the sample into the sample loop Note Make sure to flush the loop with DM water and buffer using at least 5 times the loop volume before injecting the samp
146. ns 29 0270 57 AG 6 Operations from the Instrument Display 6 4 Perform a method run 6 4 2 Quick start Step Action Graph to view the chromatogram Edit run to change any run parameters in the current run Hold to temporarily hold the run with current set flow rate valve positions and B concentration Pause to pause the current run End to end the run before it is completed Note The run begins with a default pump wash Pump wash is performed at 10 ml min for 1 min with 30 sec of buffer B wash followed by 30 sec of buffer A wash Edit run is disabled when the pump wash is in progress During pump wash the flow is directed through Wash valve to Waste Note The USB memory stick stores the result files which can be viewed using UNICORN start Also a BMP file is generated which can be viewed from any PC For details refer to Section 6 6 3 BMP result file on page 208 Do not remove the USB memory stick until the system generates the report BMP file Hold the run Follow the instruction below to hold an ongoing run Step Action 1 Run view UV absorbance 1 0 mAU Conductivity 25 0 mS cm es Flow rate 3 0 ml min f Graph Pressure 0 30 MPa Tube no 6 Edit run Hold End In the Run view tap Hold to temporarily hold the run Note Not applicable for a manual run Hold option is active only in a method run AKTA start Operating Instructions 29 0270 57 AG 177 6 Operations from the Instrum
147. ns the graphic view of the run in progress displaying the plot of the UV absorbance mAU versus Time min Saves a user method that was either created or edited Select Confirms and initiates a run from a specific tem plate or user method Create Creates a user method based on selected tem Create plate The run parameters have to be edited as needed Confirms an action Rejects an action AKTA start Operating Instructions 29 0270 57 AG 45 3 System description 3 3 Instrument Display 3 3 1 Overview of the Instrument Display Description of Instrument Display Help The KTA start Instrument Display Help is accessible from every screen on the Instrument Display by tapping the question mark in the upper right corner The Help text provides information about the content of the current screen or refers to more detailed documen tation Select the template that suits your application Affinity Chromatography AC separation based on biological recognition Desalting DS removal of low molecular weight molecules e g salt Or when changing buffer lon exchange IEX separation based on the charge of the protein Gel filtration GF separation based on the size and shape of the protein For details see AKTA TM start Operating Instructions 46 AKTA start Operating Instructions 29 0270 57 AG 3 System description 3 3 Instrument Display 3 3 2 Description of Method run 3 3 2
148. nstrument 7 KTA start Operating Instructions 29 0270 57 AG 65 4 Installation 4 3 Unpack KTA start and Frac30 Step Action 4 Hold the red strap and then lift the instrument out of the delivery box 5 Open the strap lock and remove the strap 66 KTA start Operating Instructions 29 0270 57 AG 4 Installation 4 3 Unpack KTA start and Frac30 Step Action 6 Remove the foam cushion from the top of the instrument 7 Remove the foam cushion from the bottom of the instrument by carefully lifting the instrument AKTA start Operating Instructions 29 0270 57 AG 67 4 Installation 4 3 Unpack KTA start and Frac30 Step Action 8 Remove the plastic bag by gently tilting the system back and forth while pulling out the plastic bag Unpack Frac30 Follow the instructions below to unpack the Fraction collector 68 CAUTION Take care not to damage the Dispenser arm when lifting Frac30 or when removing the plastic bag NOTICE Never lift the Frac30 by the Dispenser arm This may damage the Fraction collector Note Frac30 with packaging weighs about 6 kg No lifting equipment required one person can lift and move the Fraction collector KTA start Operating Instructions 29 0270 57 AG 4 Installation 4 3 Unpack KTA start and Frac30 Step Action i Open the Frac30 delivery box by cutting the adhesive tape at the top of the box 2 Tak
149. o return to GE may lead to delays in servicing the equipment and could be subject to additional charges 3 Visible contamination will be assumed hazardous and additional cleaning and decontamination charges will be applied No Please specify if the equipment has been in contact with any of the following Infectious or hazardous biological substances please specify a Equipment must be decontaminated prior to service return Please provide a telephone number where GE can contact you for additional information concerning the system equipment Telephone No Liquid and or gas in equipment is Water Ethanol None empty Argon Helium Nitrogen Liquid Nitrogen Other please specify I hereby confirm that the equipment specified above has been cleaned to remove any hazardous substances and that the area has been made safe and accessible Company or Name institution Position or job title Date YYYY MM DD Signed GE and GE monogram are trademarks of General Electric Company To receive a return authorization number or service number GE Healthcare Bio Sciences Corp 800 Centennial Avenue P O Box 1327 Piscataway please call local technical support or customer service NJ 08855 1327 US 2010 14 General Electric Company All rights reserved First published April 2010 DOC1149544 28 9800 27 AC 05 2014 270 AKTA start Operating Instructions 29 0270 57 AG 10 5 O
150. olume of DM wa ter buffer has completed Load the sample using the Pump The sample can be applied directly using the Pump through the Sample valve The direct sample application technique allows the application of sample volumes larger than 5 ml Follow the instructions below to apply the sample directly using the Pump Step Action 1 2 Connect a 1 mmi d ETFE tubing to port I Sample of the Sample valve Immerse the other end of the sample inlet tubing in the sample container From the Instrument Display select Method Run Method run Quick start Templates User defined Prepare system E S Choose Quick start or Templates For details see Section 6 4 Perform a method run on page 170 KTA start Operating Instructions 29 0270 57 AG 139 5 Prepare the system for a run 5 7 Sample application Step Action 4 In the Run parameters screen select the sample application via the Pump Run parameters Sample from Pump Loop Sample volume v P iml lt gt Equilibration CV volume 50 eik Wash unbound 15 0 CV volume fa 2 3 Note e When the sample is applied via the Pump the Injection valve has to be manually set to position Load default e Make sure to wash the sample inlet tubing with buffer A before immersing the tubing into the sample tube Make sure to keep sufficient volume of sample to avoid air entering the tubing e Make sure that there are
151. on connect a computer 80 connect Frac30 78 connect power 77 pump tubing 76 site requirements 60 Unpack KTA start 64 Unpack Frac30 68 Instrument display function 43 Instrument Display Create method screen 50 help 46 Method run screen 47 overview 42 Settings and service screen 51 touch buttons 43 touch screen calibration 90 L Literature 268 M Maintenance 217 219 Index cleaning before service 221 clean the instrument 231 232 disconnect column 223 long term storage 237 maintenance schedule 219 Prepare system meth ods 189 Pump wash A 126 Pump wash B 128 regular 218 replacement proce dures 230 238 safety precautions 218 short term storage 236 System cleaning 189 224 Washout fractionation tub ing 130 Manage files USB memory stick 205 Manage methods create a method 197 delete a method 204 edit a method 200 import a method 202 Manual run edit 166 end 169 pause 168 start a run 163 Manufacturing information 9 Method run hold 177 method types 171 Quick start 173 select a method 172 start a run 174 templates 179 user defined methods 187 N Nomenclature conventions 6 Notes and tips 8 O Operations checklist 157 cold room 151 from the instrument display Manage methods 195 perform a manual run 162 perform a method run 170 from UNICORN start 210 monitor the run 165 workflow 156 P Power off 1
152. on Colector seserinis aur araka betretetoe 255 8 7 Storage OT TS WS I sssrinin EAE E ENESE 236 KTA start Operating Instructions 29 0270 57 AG 3 Table of Contents 10 11 8 8 Replacement of tubing and filters xvccctessstcssccscucctutuvnrsctesentsiadecbtvensiiesnetanstgetCawaddeceeadbtannttactausbtos 238 8 8 1 Replace the inlet filters ss ssssssssssssssssssrrirsssssssssssssssrirsrrrrsssssssssssrssrrrrsrrressssssnsssnrrrrrrrrrrrs 239 8 8 2 Replace the tubing and CONMCCIONS cnsununinaidunuanaandmnnnmmeamanannas 240 Troubleshotin sicastosrcrsssrsscarsicvertoranpiaeeatreteeaieieemucnnennnans 242 9 1 Introduction to troubleshooting ssssssssssssssssssssssssssssssssssseessssrrssrierrirrrerrerensrennsnssnnsnnnsnnnnnnnnsnnnnnnns 243 D PAA TOE 01616 10 E E A E ET 245 9 3 System error report escssssccssessessessececossonssesnccucoscoseassacsucsscaseassassanssessessessussseseeaseassassaccucoseasenssnesacess 256 Reference information esseseeseeserseesesseseeseesersessesseseeseesersessesseseeseesersesseeseseseso 258 WUE Pe O a e E E E E NE 259 il 9 ummm E EE E S E EE E E 265 Wo Er E ee EE 268 10 4 Health and Safety Declaration FOTN scccpstactssscceitetsnrcincssiaunsencasrdcecseenecoeastiadnteationtnes 269 105 Ordening Mormaii en E RES 271 19 o 1 4 5 Seen Re E ona Rea Me ene EE EE Me trons Tan ante E mae ee 275 Te System Perommance REDONE suissa enni EE ee tenessces ieee estates 276 E E AA MIEN E AAE EE E R
153. ons 5 3 1 Calibration guide 5 501 Calibration guide When to calibrate Note The instrument is pre calibrated at delivery therefore no calibration is required when the instrument is installed However if the System performance test fails it is recommended to calibrate the modules The table below provides recommendations for when modules should be calibrated Module When to calibrate Instrument Display If there are any issues with the response of the touch screen Pressure sensor If the pressure is outside the range of 0 03 MPa perform Zero offset Pump When chromatography run conditions are changed e g viscosity of sample or buffer temperature back pressure Pump and pump tubing requires calibration regularly Recommended once a week After the pump tubing has been replaced with new tubing Note Do not leave the pump tubing inside the Pump when the Pump is not running UV Monitor When the signal is unstable or readings appear to be incorrect After cleaning or after replacing the UV flow cell When error warning is seen on power ON When baselining is ignored when the UV flow cell is clean Before and after performing runs in the cold room Conductivity Monitor When the signal is unstable or readings appear to be incorrect After replacing the Conductivity flow cell AKTA start Operating Instructions 29 0270 57 AG 89 5 Prepare the system for a run 5 3 Calibrations 5
154. ons 29 0270 57 AG 19 2 Safety instructions 2 1 Safety precautions NOTICE To avoid damage to KTA start or other equipment when installing or moving the instrument follow the instructions below e Vents on KTA start To ensure adequate ventilation keep papers and other objects away from the vents of the instru ment e Any computer used with the equipment shall comply with IEC 60950 and be installed and used according to the manufactur er s instructions e Frac30 should not be connected or disconnected from KTA start when the instrument is powered ON System operation WARNING To avoid personal injury when operating AKTA start follow the in structions below e Rotating the instrument Make sure that there is always at least 20 cm of free space around AKTA start to allow for suffi cient ventilation When turning or moving the instrument take care not to stretch or squeeze tubing or cables A disconnected cable may cause power interruption or network interruption Stretched tubing may cause bottles to fall resulting in liquid spillage and shattered glass Squeezed tubing may cause in crease in pressure or block liquid flow To avoid the risk of knocking over bottles always place bottles on the buffer tray and turn or move carefully e Hazardous chemicals during run When using hazardous chemicals run the System cleaning template to clean and flush the entire system tubing with distilled w
155. or a run 5 3 Calibrations 5 3 4 Pump calibration Step Action 5 In the Pump screen tap Calibration Result The Calibration screen opens Calibration 2 Flow Rate gi 0 5 ml min Start flow Collected f _ iy 1 2 ml Calibrate Volume L Back A 6 In the Calibration screen set the intended flow rate in the range 0 5 to 5 ml min Use the up down arrows to set the value or use numpad to type in the value 7 In the Calibration screen tap Start flow to start the Pump Result The Pump starts and the pumped DM water will be collected in the pre weighed collection tube Collect water for at least one minute in the pre weighed collection tube 8 Estimate accurately the volume of the water collected in the pre weighed collection tube then set the Collected volume value equal to this volume 9 In the Calibration screen tap Calibrate then confirm the action OK to carry out the calibration 10 Verify that the Pump was correctly calibrated Repeat the steps 6 and 7 to confirm that the volume of the collected water corresponds to the intended flow rate Note If the collected water does not correspond to the required volume i e the Pump does not deliver the liquid with the intended flow rate inspect the condition of the pump tubing Replace the pump tubing if necessary and then perform re calibration AKTA start Operating Instructions 29 0270 57 AG 95 5 Prepare the system for a run 5 3 Calib
156. or many purification schemes it is important to collect fractions of the eluent AKTA start offers the fractionation options presented in the table below Instrument configuration Fractionation options KTA start UNICORN start Frac30 Fixed volume fractionation Peak fractionation Level based Slope based KTA start UNICORN start Single Peak collection Level based KTA start Frac30 Fixed volume fractionation KTA start e Collection of elution volume Handling the delay volume For setting the delay volume refer to Section 5 8 Prepare the Fraction collector on page 147 Fractionation with Frac30 The delay volume is collected in the first tube T1 followed by the rest of the fractions in the subsequent tubes Fractionation without Frac30 The delay volume is collected in the collection container along with the fraction i e Total collected volume delay volume fraction volume KTA start Operating Instructions 29 0270 57 AG 159 6 Operations from the Instrument Display 6 2 Fractionation Fractionation using AKTA start For detailed instructions on fractionation operations using UNICORN start refer to UNICORN start 1 0 User Manual AKTA start with Frac30 A representative chromatogram depicting fractionation using fixed volume fraction collection in AKTA start with Frac30 is shown below Note Make sure to set fraction volumes that suit the column being used and use an adequate
157. or national regulations for safe operation maintenance and decommissioning of the equip ment 18 AKTA start Operating Instructions 29 0270 57 AG 2 Safety instructions 2 1 Safety precautions Installing and moving the instrument WARNING To avoid damage to person when installing or moving AKTA start follow the instructions below e Moving the instrument horizontally One person is recommend ed when moving the instrument horizontally e Supply voltage Make sure that the supply voltage at the wall outlet corresponds to the marking on the instrument before connecting the power cord e Power cord Only use grounded power cords delivered or ap proved by GE e Access to power switch and power cord with plug Do not block access to the power switch and power cord The power switch must always be easy to access The power cord with plug must always be easy to disconnect e Installing the computer optional The computer should be installed and used according to the instructions provided by the manufacturer of the computer e Disconnect power Always switch off power to KTA start be fore an instrument module is removed or installed or a cable is connected or disconnected CAUTION To avoid damage to person when installing or moving AKTA start follow the instructions below Protective ground AKTA start must always be connected to a grounded power outlet AKTA start Operating Instructi
158. p Action 1 To remove the test tube holder unsnap the snap locks one by one and se quentially for ease of removal 2 To fit the test tube holder on the Bowl base e Align the holder with the help of the aligning groove e Snap fit the part by operating one snap at a time in a continuous se quence by pressing at the edge of the part KTA start Operating Instructions 29 0270 57 AG 235 8 Maintenance 8 7 Storage of the instrument 8 7 Storage of the instrument Required material The following materials are required e DM water e 20 ethanol e 0 75 mm i d PEEK tubing e Waste container Short term storage If the system is not going to be used for a few days follow the instructions below to prepare the system for short term storage Note Before cleaning the flow path remove the column and re connect the flow path See Section 8 3 1 Disconnect the column on page 223 Step Action 1 Immerse the buffer and the sample inlet tubing in DM water 2 In the KTA start home screen tap Manual run For detailed instructions see Section 6 3 Perform a manual run on page 162 Set the run parameters according to the table below Parameter Setting Flow rate 1 ml min Pressure limit 0 5 MPa Select Buffer Sample Buffer Wash valve Column Outlet valve Waste 3 Tap Run to start the manual run Pump 20 ml of DM water through the system 236 KTA start Operating Instructions 29 0270 57
159. pa radio button to select a template that suits your application e To continue with the selected technique tap Select Run parameters Column volume ml Flow rate ml min p S Pressure limit MPa Toa us Q EE AKTA start Operating Instructions 29 0270 57 AG 6 Operations from the Instrument Display 6 4 Perform a method run 6 4 3 Templates Step Action e Set the run parameters Column volume column volume ml Flow rate flow rate ml min Pressure limit pessure limit MPa e Tick the checkbox Save Result to USB if you want to save the result The result file name can be edited by setting the digits in the range 00 to 99 e Tap the forward arrow to access additional run parameters Note Make sure that the values for the Column volume Flow rate and Pressure limit are appropriate for the chosen column Refer to the column manual for details If the pressure reaches above the set limit the instrument will enter the Pause state Note The USB memory stick stores the result files which can be viewed using UNICORN start Also a BMP file is generated which can be viewed from any PC For details refer to Section 6 6 3 BMP result file on page 208 Do not remove the USB memory stick until the system generates the report BMP file Run parameters Sample from Pump Loop Sample volume v 1 A ml Equilibration volume Wash unbound volume vom CV CV 2 3 e Select the mode of s
160. persistent clean the UV flow cell See Section 8 4 Cleaning the UV flow cell on page 227 Buffer is impure Check if the signal is still noisy with DM water Air dissolved in the buffer or air bub bles seen in the buffer inlet tub ings UV flow cell not clean Degas the buffer before use Use a tech nique available in the laboratory such as vacuum degassing or sonication Perform Pump wash to remove the air bubbles from the flow path Clean the UV flow cell See Section 8 4 Cleaning the UV flow cell on page 227 UV flow cell not mounted properly locknut not tight en Remove the protective cover and check the flow cell Tighten the locknut Ghost peaks Air dissolved in the buffer Degas the buffer before use Use a tech nique available in the laboratory such as vacuum degassing or sonication Flow path not clean Clean the system flow path See Sec tion 8 3 Cleaning the system flow path on page 222 Column not clean Clean the column according to the col umn instructions Low sensitivity UV flow cell not clean Clean the UV flow cell See Section 8 4 Cleaning the UV flow cell on page 227 Waves on the gra dient Switch valve tim ing not optimized Perform Switch valve timing to optimize the switch valve timing See Section 5 4 4 Switch valve timing on page 117 AKTA start Operating Instructions 29 0270 57 AG 9 Troubleshooting 9
161. ponse to B changes KTA start Operating Instructions 29 0270 57 AG Buffer valve mal function See Buffer valve troubleshooting Irregular flow Tubing not clean Check that the Pump functions properly See Pump troubleshooting Make sure that the tubing is washed properly Run System cleaning to clean the system flow path See Section 8 3 Cleaning the system flow path on page 222 249 9 Troubleshooting 9 2 Basic troubleshooting Possible cause Action Air bubbles in the flow path Worn out pump tubing Remove the air bubbles trapped in the flow path by flushing the flow path with DM water or buffer according to the pro cedure below e remove the column and re connect the flow path see Section 8 3 1 Dis connect the column on page 223 e perform a manual run with a flow rate of 5 ml min for around 10 min utes e observe the graph until the pulsations are no more visible and the curve Is stable Note If there is air trapped in the flow path the flow is not accurate and pulsations can occur affecting the output signals Replace the pump tubing See Sec tion 8 8 2 Replace the tubing and connec tors on page 240 Pump not calibrat ed Calibrate the Pump See Section 5 3 Cali brations on page 88 Pump tubing not properly posi tioned Place the pump tubing in the pump hood maintaining equal distance on both the sides Worn out pump tubing
162. power cord AKTA start Operating Instructions 29 0270 57 AG 77 4 Installation 4 5 Install KTA start Step Action 2 Connect the power cord to the Power input connector on the left side of the instrument and to a grounded wall outlet 100 240 VAC 50 60 Hz CAUTION AKTA start is filled with denaturated alcohol 50 C2H50OH ethanol at delivery The denatured alcohol mixture can be hazardous to humans if consumed Flush out the denaturated alcohol before assembling testing or integrating AKTA start into the intended process context Note For instructions on how to clean the flow path see Section 8 3 Cleaning the system flow path on page 222 Connect Frac30 to AKTA start NOTICE Frac30 should not be connected or disconnected from KTA start when the instrument is powered ON Follow the instructions below to connect Frac30 to KTA start 78 KTA start Operating Instructions 29 0270 57 AG 4 Installation 4 5 Install AKTA start Step Action i Connect the Frac30 cable between the intended ports on the back of the Fraction collector and the instrument Note The supply voltage for Frac30 is distributed from KTA start When the Frac30 cable is connected the screws attached to the connector should be tightened 2 Switch on AKTA start 3 Enable the connection of Frac30 from the Instrument Display e inthe KTA start home screen tap Settings and service Result
163. puter Check the content of the system report files INSTLOG TXT and ERRORLOG TXT The content should look similar to the images below a EE alle Nea Fle Edk Format View Help AK TA start ser alno 1234567891 1234567895 Firmwere Version i V2 01 19 501 Over Pressure FPGA Version v2 00 00 301 Frac Module Failure ep ae Bethe 603 Meeel operation Restart hstrument UV LED run tine 63 2Hrs 301 Frec Moduls Failure Buffer valve count 1 Wash walve count 9 Outlet valve count 5 emple vahe count 25 4 Use the system report in further contacts with GE Service Engineer KTA start Operating Instructions 29 0270 57 AG 257 10 Reference information 10 Reference information About this chapter This chapter lists the technical specifications of AKTA start The chapter also includes a list of wetted materials a chemical resistance guide Health and Declaration form for service and ordering information In this chapter This chapter contains the following sections Section See page 10 1 Specifications 259 10 2 Chemical resistance 265 10 3 Literature 268 10 4 Health and Safety Declaration Form 269 10 5 Ordering information eT 258 AKTA start Operating Instructions 29 0270 57 AG 10 Reference information 10 1 Specifications 10 1 Specifications Introduction This section lists the specification data of AKTA start For component data see the AKTA start Maintenance Manual System specifications Parameter Syste
164. quirement in SJ T11363 2006 x Indicates that this toxic or hazardous substance contained in at least one of the homoge neous materials used for this part is above the limit requirement in SJ T11363 2006 Data listed in the table represents best information available at the time of publication 32 KTA start Operating Instructions 29 0270 57 AG 3 System description 3 System description About this chapter This chapter provides an overview of AKTA start the Instrument Display that allows the user to operate and control the system and Frac30 Fraction collector In this chapter This chapter contains the following sections Section See page 3 1 System overview 34 3 2 Instrument 38 3 3 Instrument Display 41 AKTA start Operating Instructions 29 0270 57 AG 33 3 System description 3 1 System overview 3 1 System overview Introduction AKTA start is the main instrument included in a liquid chromatography system intended primarily for preparative purification of proteins at laboratory scale The system can be used for a variety of research purposes to fulfill the needs of the users in the academia and in life sciences industry AKTA start is operated and controlled from the Instrument Display In addition the UNICORN start software can be used to control AKTA start and to analyze the data ac quired during chromatography runs UNICORN start offers several additional features that are described in detail in UNI
165. r respective owners Any use of UNICORN start is subject to GE Healthcare Standard Software End User License Agreement for Life Sciences Software Products A copy of this Standard Software End User License Agreement is available on request UNICORN start 2013 2015 General Electric Company 2013 2015 General Electric Company All rights reserved First published Sept 2013 All goods and services are sold subject to the terms and conditions of sale of the company within GE Healthcare which supplies them A copy of these terms and conditions is available on request Contact your local GE Healthcare repre sentative for the most current information GE Healthcare Europe GmbH Munzinger Strasse 5 D 79111 Freiburg Germany GE Healthcare UK Limited Amersham Place Little Chalfont Buckinghamshire HP7 9NA UK GE Healthcare Bio Sciences Corp 800 Centennial Avenue P O Box 1327 Piscataway NJ 08855 1327 USA GE Healthcare Japan Corporation Sanken Bldg 3 25 1 Hyakunincho Shinjuku ku Tokyo 169 0073 Japan 29 0270 57 AG 02 2015 a4
166. rate e A suitable sample application technique will be used See Section 5 7 Sample appli cation on page 135 e The buffer inlet tubing is immersed into correct buffer bottles Consider the volume required for the intended application e The waste tubing is inserted into an appropriate waste container Consider container size and Its material e No tubing is twisted or blocked and the flow path is free from leakage AKTA start Operating Instructions 29 0270 57 AG 157 6 Operations from the Instrument Display 6 1 Introduction The Fraction collector configuration needs to be either enabled or disabled as re quired If the Fraction collector is used make sure to use tubes with the same tube size The delay volume is set Specifications of run parameters Parameter Flow rate 0 5 to 5 0 ml min Increment Column volume CV 1 to 1000 ml O to 150 ml Pressure limit 0 1 to 5 bar Sample volume Pump 0 1 to 1000 0 ml 0 1 to 1000 ml Sample loop 0 1 to 1000 ml Superloop 0 1 to 1000 ml Wash volume 0 0 to 50 0 CV Equilibration volume 0 0 to 50 0 CV Elution volume 0 0 to 100 CV Target B concentration O to 100 Gradient length 0 0 to 100 0 Fractionation volume 0 5to15 ml AKTA start Operating Instructions 29 0270 57 AG 6 Operations from the Instrument Display 6 2 Fractionation 6 2 Fractionation Fractionation options F
167. rations 5 3 5 UV Monitor calibration 5 3 5 UV Monitor calibration Set the Cell path length If the UV flow cell was replaced a new Cell path length value has to be set Note Make sure that the UV flow cell is assembled tightly and the inlet and outlet tubing is fitted tightly to the UV Monitor No stray light should enter the UV Monitor Make sure that there are no air bubbles in the UV flow cell Prerequisites e Buffer A DM water e Buffer B freshly prepared 1 acetone solution which is expected to give an Abs value of 340 MAU Follow the instructions below to set the Cell path length Step Action 1 In the KTA start home screen tap Settings and service to access the in strument modules then tap Next Next Result The Settings and service Screen 3 opens Settings and service 3 3 Back ED 2 In the Settings and service screen tap UV to access the UV Monitor options Result The UV screen opens UV 2 Flow cell path length Back or 96 AKTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 3 Calibrations 5 3 5 UV Monitor calibration Step Action 3 In the UV screen tap Flow cell path length to access the flow cell options For a description of the UV Monitor options refer to the AKTA start Maintenance Manual Result The Flow Cell Path Length screen opens Flow cell path length Abs o0 mau Set baseline Back tat In the Flow Cell Path Le
168. rdering information For ordering information visit www gelifesciences com AKTA Accessories list 10 Reference information 10 5 Ordering information Part Accessory description Code no Marprene Tubing 29 0240 12 Peristaltic Pump 29 0239 92 Solenoid valve Buffer valve 29 0238 95 Sample valve 29 0238 96 Wash valve 29 0238 97 Outlet valve 29 0238 98 Manual Injection valve Injection valve Manual 29 0239 58 Valve kit Manual INV 29 0239 17 Mixer Mixer AKTA start 29 0239 60 UV UV module AKTA start 29 0240 18 Flow Cell 2 mm UPC 900 29 0113 25 Conductivity Conductivity Cell AKTA start 29 0240 21 Sample loops Sample Loop PEEK 10 ul 18 1120 39 Sample Loop 100 ul INV 907 18 1113 98 Sample Loop 500 ul INV 907 18 1113 99 Sample Loop 1 0 ml INV 907 18 1114 01 Sample Loop 2 0 ml INV 907 18 1114 02 Sample Loop 5 ml PEEK 18 1140 53 Sample Loop 18 1161 24 Superloop Superloop 10 ml KTA 18 1113 81 Superloop 50 ml AKTA 18 1113 82 Superloop 150 ml 18 1023 85 AKTA start Operating Instructions 29 0270 57 AG 271 10 Reference information 10 5 Ordering information Part Accessory description Code no Fittings Tubing Connector 1 8 18 1121 17 Ferrule for 1 8 tubing 18 1121 18 Union Luer Female HPLC Male 18 1112 51 Fingertight Connector 1 16 18 1112 55 Stop plug 1 16 PKG 5 18 1112 52 Stop plug 5 16 PKG 5 18 1112 50 Union
169. rge Gel filtration columns are used it is recommended to pre equilibrate the column before starting the run Run parameters Sample from Pump Sample volume v 50 ym mi O bu lU ACO1 to USB e The mode of sample application is Pump default Note Sample application using Pump is used for all Quick start methods to automate sample loading or to have an unattended chromatographic run Note Sample application via Loop is not applicable AKTA start Operating Instructions 29 0270 57 AG 175 6 Operations from the Instrument Display 6 4 Perform a method run 6 4 2 Quick start 176 Step Action e Enter the sample volume in the Sample volume field Use up down arrows to set the values or use numpad to type in the values e Tick the checkbox Save Result to USB if you want to save the result The result file name can be edited by setting the digits in the range 00 to 99 e Tap Run to start the run Result The Run view will be displayed Note Other run parameters can be edited using the Edit run option available in the Run view screen 4 Run view UV absorbance 1 0 mAU Conductivity 25 0 mS cm x Flow rate 3 0 ml min Graph Pressure 0 30 MPa Tube no 6 Edit run End In the Run view screen the following options are available to monitor and control the ongoing run for details see Section 6 3 2 Monitor and control the run on page 165 AKTA start Operating Instructio
170. rnal parts may be worn Replace valve Refer to KTA start Maintenance Manual High back pres sure Flow path not clean Clean the system flow path See Sec tion 8 3 Cleaning the system flow path on page 222 Unable to load sample to the loop Manual injection valve is in the In ject position Turn the Injection valve to Load position while loading the sample to the loop The Valve or the loop is blocked Clean the system flow path If problem persists replace the valve loop AKTA start Operating Instructions 29 0270 57 AG 9 Troubleshooting 9 2 Basic troubleshooting Pressure sensor Description Possible cause Action Error 501 Over Blockage in Check the tubing and valves by dis pressure tubing valves connecting one at a time Clean or or in the col replace when a blockage is found umn Clean the column separation media Improperly se with suitable solution 1 M NaOH or lected flow replace the column with a new one rate Check the specification of the columns for correct flow rate Pressure sensor is Pressure sensor Contact a GE Service Engineer not functioning failure or failure of Pressure Curve Is component in not displayed on Main board the screen User Information Messages Description Possible cause Instruction ignored message Instruction This message is shown when it is not ignored not allowed to set the inlets possible to give the in
171. rt Operating Instructions 29 0270 57 AG 91 5 Prepare the system for a run 5 3 Calibrations 5 3 3 Pressure sensor zero offset oe Zero offset 92 Pressure sensor zero offset Follow the instructions below to set the pressure as zero when the Pressure sensor is exposed to atmospheric pressure only Zero offset Note Step Pressure Calibration is performed by a GE Service Engineer only after the Pressure sensor is replaced Action l Disconnect the inlet tubing from the Pressure sensor in order to expose the sensor to atmospheric pressure only Note Make sure that the Pump is OFF before disconnecting the tubing In the KTA start home screen tap Settings and service to access the in strument modules Result The Service and settings screen opens Settings and service Fraction collector Pressure sensor 1 3 E S Next In the Settings and service screen tap Pressure sensor to access the Pressure sensor options Result The Pressure sensor screen opens Pressure sensor User operations P set MPa Zero offset GEHC service only Stop flow Calibrate Back E A KTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 3 Calibrations 5 3 3 Pressure sensor zero offset Step Action 4 In the Pressure sensor screen tap Zero offset Result A Message screen opens Message A Make sure No Pre
172. rument Display 6 4 Perform a method run 6 4 3 Templates Step Action e Configure the run parameters for Elution Option set as Gradient Bound proteins are eluted with continuous change of buffer B composition to increase eluent strength over specified time Target conc B maximum buffer B concentration level to be set in the gradient Gradient volume volume needed to elute the bound protein from the column Fractionation volume volume of the fraction to be collected when the Fraction collector is enabled e Tap back arrow to view or edit run parameters e Tap Run to start the run Result The Run view screen will be displayed Run view UV absorbance 1 0 mA amp AU Conductivity 25 0 mS cm Flow rate 3 0 ml min Graph Pressure 0 30 MPa Tube no 6 Edit run End Note The run begins with a default pump wash Pump wash is performed at 10 ml min for 1 min with 30 sec of buffer B wash followed by 30 sec of buffer A wash Edit run is disabled when the pump wash is in progress During pump wash the flow is directed through Wash valve to Waste Gel filtration Desalting Buffer Exchange Follow the instruction below to start a run based on Gel Filtration or Desalting template Note Make sure that a USB memory stick is connected to the instrument If the instrument does not detect a USB memory stick the result will not be saved Note Before starting the run set the status of the Fract
173. run For detailed instructions on cleaning the system using the System cleaning template see Section 8 3 2 System cleaning on page 224 Step Action L Method run Quick start Templates User defined Prepare system In the Method run screen tap Prepare system to access the system methods AKTA start Operating Instructions 29 0270 57 AG 189 6 Operations from the Instrument Display 6 4 Perform a method run 6 4 5 Prepare system methods Step Action 2 ry Select system method System cleaning System performance method In the Select system method screen e Select System cleaning e Tocontinue with the System cleaning method tap Select Please follow instructions 1 Disconnect the column 2 Connect injection valve to UV with suitable tubing and connectors 3 Place the buffer tubings A amp B in cleaning solution When completed press Continue e Perform the operations described on the display 1 Disconnect the column 2 Connect Injection valve to UV with suitable tubing and connectors 3 Place the buffer tubings A amp B in cleaning solution For detailed instructions see Section 8 3 2 System cleaning on page 224 e Tap Continue to start the System cleaning run 190 AKTA start Operating Instructions 29 0270 57 AG 6 Operations from the Instrument Display 6 4 Perform a method run 6 4 5 Prepare system methods Step Action 4 System cleaning in progress UV
174. s to be set AKTA start Operating Instructions 29 0270 57 AG 163 6 Operations from the Instrument Display 6 3 Perform a manual run 6 3 1 Manual run Step Action Note Make sure that the values for the Flow rate and Pressure limit are appropriate for the chosen column Refer to the column manual for details If the pressure reaches above the set limit the instrument will enter the Pause state Manual run lt l Outlet valve Collection une Bo B dian M os O a2 e Toggle to set which valve positions are open Sample valve set as Buffer or Sample so that the flow is delivered from either the buffer inlets or the sample inlet Washvalve set as Column or Waste to direct the flow to either the column or to waste Outlet valve set as Collection or Waste to direct the flow to either the Fraction collector or to waste e Setthe Fractionation volume the volume of the fraction to be collected when the Fraction collector is enabled Use up down arrows to set the value or use numpad to type in the value Note To collect fractions make sure that the Fraction collector is enabled If the Fraction collector is enabled place the required number of tubes of adequate volume in the Bow assembly If enabled the Fraction collector will home to position 1 at the beginning of every run e Tap Run to start the run Result The Run view screen will be displayed 164 AKTA start Operating Instr
175. sed Follow the instructions below to set the delay volume Step Action 1 In the AKTA start home screen tap Settings and service to access the in strument modules Result The Settings and service Screen 1 opens Settings and service Fraction collector Pressure sensor System Q Next 2 In the Settings and service screen tap System to access the system options Result The System screen opens System 2 Serial 00000000000000000001 igi pn ie Firmware update setting Switch valve timing PESE Use AKTA start Operating Instructions 29 0270 57 AG 149 5 Prepare the system for a run 5 8 Prepare the Fraction collector 150 Action In the System screen tap Delay volume setting to access the settings Result The Delay volume settings screen opens Delay volume setting Outlet valve to Fraction collector Tube ID mm Tubelength mm TE ca Total Delay volume UV to Fraction collector 0 49 ml Back A Enter the internal diameter ID in the Tube ID field and length of the tubing from the Outlet valve to Fraction collector in the Tube length fields and then tap Save Result The total delay volume from UV to Fraction collector is displayed in the Total delay volume field Note The delay volume from UV to Outlet valve is constant 0 27 ml in all KTA start instruments if the recommended tubing lengths and i d are used Note
176. sistant to the substances used Follow local and or national regulations for safe operation maintenance and decommissioning of the equipment Tip If hazardous chemicals are used for system or column cleaning wash the system or columns with a neutral solution in the last phase or step In this section This section contains the following subsections Section See page 8 3 1 Disconnect the column 225 8 3 2 System cleaning 224 222 AKTA start Operating Instructions 29 0270 57 AG 8 Maintenance 8 3 Cleaning the system flow path 8 3 1 Disconnect the column 8 3 1 Disconnect the column Introduction The column should be removed from the flow path before cleaning the system flow path The flow path has to be re connected between the manual Injection valve port 1 and the UV inlet For column cleaning procedures and storage instructions refer to the column catalogue Instruction Follow the instructions below to remove the column and re connect the flow path Step Action 1 Disconnect the tubing from the column as indicated by the arrows in the illustration below 1 2 Re connect the flow path between the Injection valve and the UV Monitor as indicated by the arrows in the illustration below 2 Join the tubing by using the union mounted on the tubing connected to the UV flow cell AKTA start Operating Instructions 29 0270 57 AG 223 8 Maintenance 8 3 Cleaning the system flow path 8 3 2 System c
177. ssure in the system Co ico 5 Tap OK if there is no back pressure in the system i e the inlet tubing from the Pump was disconnected from the Pressure sensor Result The atmospheric pressure is set as zero 6 Re connect the inlet tubing from Pump to the Pressure sensor AKTA start Operating Instructions 29 0270 57 AG 93 5 Prepare the system for a run 5 3 Calibrations 5 3 4 Pump calibration 5 3 4 Instruction 94 Pump calibration Follow the instructions below to calibrate the Pump Step iL 2 Action Immerse the buffer inlet tubing A in demineralized DM water Place the outlet tubing from the Wash valve in a pre weighed collection tube Note Before starting the calibration prime the flow path with water and make sure that the outlet tubing where the pumped water is collected is filled with DM water This ensures that the volume of the collected water corresponds to the pumped volume In the AKTA start home screen tap Settings and service to access the in strument modules Result The Settings and service Screen 1 opens Settings and service Fraction collector Pressure sensor Pump System 1 3 Q Next In the Settings and service screen tap Pump to access the Pump options Result The Pump screen opens Pump 2 Calibration Pump tubing log Back Q AKTA start Operating Instructions 29 0270 57 AG 5 Prepare the system f
178. stored on a USB memory stick connected to the instrument When Create method is selected further options are displayed in the Create method screen Create method Delete USB import The operations available under Create method are briefly described below Description Create Displays the method templates that can be used to create a new method Edit Displays the user methods stored on the instrument that can be edited if required USB Import Displays a list of user methods stored on a USB memory stick that can be imported to the instrument Delete Displays the user methods stored on the instrument that can be deleted if required 50 AKTA start Operating Instructions 29 0270 57 AG 3 System description 3 3 Instrument Display 3 3 4 Description of Settings and service 3 3 4 Description of Settings and service Settings and service options The display option Settings and service allows the user to perform maintenance calibration setting the delay volume diagnostics and troubleshooting of the modules located on the wet side of the instrument For a brief description of the modules see Section 3 2 Instrument on page 38 Detailed instructions for calibrating the Pump the monitors and the Instrument Display are presented in Section 5 3 Calibrations on page 88 The options available under Settings and service are listed in the table below Function Settings and service Screen 1
179. struction during during gradient Gradient Occurrence message The instruction This message appears after the delay Outlet valve is not possible to issue dur volume has passed and no fractionation ing an active fractionation is active Occurrence message Instruction ignored Thismessage appears when the instruc Stop fractionation is only allowed during tion Fractionation has not been execut fractionation ed the Stop fractionation instruction shall be ignored and an occurrence shall be issued Warning message Instruction ignored This warning message appears when Peak fractionation is not allowed during an instruction is not possible to execute fractionation or Single peak collection during Single peak collection or during Fractionation AKTA start Operating Instructions 29 0270 57 AG 253 9 Troubleshooting 9 2 Basic troubleshooting 254 Description Warning message Last tube has been reached change tubes in the fraction collector and press Continue to continue the run with fractionation Press Cancel fractionation to continue the run without fractionation flow is diverted to Flow through Waste position Possible cause This message appears when the last tube has been reached Occurrence message Last tube has been reached and the run has continued without fractionation This message appears if Continue without fractionation is selected Warning message Instruction ignored Single p
180. t Step Action 5 Tap Exit to close the screen when the System performance method is completed Run ended 2 UV absorbance 1 0 mAU Conductivity 25 0 mS cm Flow rate 0 0 ml min Jom Pressure 0 30 MPa Tube no 6 Exit Note Make sure not to unplug the USB memory stick until the system generates the report BMP file 6 Review the report whether the test passed or failed based on the Acceptance criteria presented below Acceptance criteria Time Activity Check Approved interval min 0 Pump wash Wash valve posi Mobile phase out tion through Waste 1 1 ml min 0 B flow through Back pressure lt 0 05 MPa Outlet valve Waste position 2 Repeat UV Auto zero 3 5 ml min Back pressure 0 06 to 0 2 MPa UV level 10 MAU Conductivity level 1mS cm 4 1 ml min Sample valve Sam Max UV level 300 to 380 MAU ple position Max Conductivity 65 to 95 mS cm level 7 1 ml min Sample valve Buffer position KTA start Operating Instructions 29 0270 57 AG 111 5 Prepare the system for a run 5 4 System performance 5 4 2 System performance method from AKTA start 112 Activity Request switch Injection valve to Inject position Max UV level Max Conductivity level Approved interval 300 to 380 mAU 65 to 95 mS cm Request switch Injection valve back to Load position Start gradient O to 100 B in 10 minutes start fractiona tion collection
181. t Description Screen name area Information area The example shows the KTA start home screen 42 Help button KTA start Operating Instructions 29 0270 57 AG Instrument software 3 System description 3 3 Instrument Display 3 3 1 Overview of the Instrument Display AKTA start offers the following functionality menu displayed on the AKTA start home screen as presented in the table below For detailed workflows on the different options see Chapter 6 Operations from the Instrument Display on page 155 Option Method run Manual run Description Use a quick start or a method template to perform a run Perform arun by providing parameters manually Create method Create edit import and delete user methods Settings and service Description of buttons on the Instrument Display Configure settings and calibrate modules perform diagnostics tests The Instrument Display includes the following touch buttons Button Description Opens a new dialog screen providing information about the content of the current screen or indi cates where further information or instructions can be found Opens the KTA start home screen Forward Return Opens the next screen in the current workflow Opens the previous screen in the current workflow KTA start Operating Instructions 29 0270 57 AG 43 3 System description 3 3 Instrument Display 3 3 1 Overview of th
182. t Lo J Run log To see the number of hours the Frac drive has been active and to reset the number of hours of drive usage Pressure sensor Zero offset Allows to set the pres sure to zero The Pump has to be off cameras 2 and the flow path connection to the User operations wo E es Pressure sensor has to ee open P set Shows the pressure in the flow GEHC service only path Stop flow Calibrate Note Back ED Pressure Calibration is performed by a GE Service Engineer only after the Pressure sensor is replaced 52 KTA start Operating Instructions 29 0270 57 AG 3 System description 3 3 Instrument Display 3 3 4 Description of Settings and service Mole UI Function Pump Calibration To calibrate the Pump so that it delivers liquid with an accu rate flow rate Pump 2 e Tonote the number of hours the Diagnostics pump tubing has been in use Tubing run and to reset the Ay cock D cy count of tubing usage after the pump tubing is replaced e Tonote the number of hours the Pump has been in use Pump run and to reset the count of Pump usage after the Pump is replaced Diagnostics To adjust the flow rate and start stop the flow Pump AKTA start Operating Instructions 29 0270 57 AG 53 3 System description 3 3 Instrument Display 3 3 4 Description of Settings and service Module System System System Ey Serial 00000000000000
183. t conc B maximum buffer B concentration level to be set in the gradient Gradient volume volume needed to elute the bound protein from the column Fractionation volume volume of the fraction to be collected when the Fraction collector is enabled e Tap Save to save the new method Result A Message screen that requires to confirm the action will be dis played AKTA start Operating Instructions 29 0270 57 AG 199 6 Operations from the Instrument Display 6 6 Manage methods and files 6 6 1 Create method Step Action 6 Message A Are you sure you want to save the method Yes No Tap Yes to confirm the saving of the method or tap No to cancel the action and return to set the run parameters Note The created method will be saved under the User defined methods menu The system can store up to 10 methods only Delete existing methods if new methods need to be stored Edit a method Follow the instructions below to edit a user defined method Step Action l 200 B Create method Create In the Create method screen tap Edit to access the methods Result The Select method to edit screen opens AKTA start Operating Instructions 29 0270 57 AG 6 Operations from the Instrument Display 6 6 Manage methods and files 6 6 1 Create method Step Action 2 Select method to edit 2 Method 1 O Method 2 O Method 3 Method 4 Back E3 Ede e Inthe
184. t is used to direct the flow to the Fraction collector or to Waste 40 KTA start Operating Instructions 29 0270 57 AG 3 System description 3 3 Instrument Display 3 3 Instrument Display Introduction This section provides a description of the AKTA start Instrument Display and the functions that are accessible from the display In this section This section contains the following sub sections Section See page 3 3 1 Overview of the Instrument Display 42 3 3 2 Description of Method run 47 3 3 3 Description of Create method 50 3 3 4 Description of Settings and service 51 KTA start Operating Instructions 29 0270 57 AG 41 3 System description 3 3 Instrument Display 3 3 1 Overview of the Instrument Display 3 3 1 Overview of the Instrument Display Introduction The Instrument Display is located on the front side of AKTA start The Instrument Display enables the user to control the system by selecting the intended operation Start a run and control an ongoing run View the progress of the ongoing run Manage user defined methods Perform maintenance and service This section provides a brief description of the functions of the Instrument Display Note Do not operate the Instrument Display with a sharp and hard object Illustration of the Instrument Display The illustration below shows the location and detailed view of the Instrument Display AKTA start Method run Create method Par
185. tem settings chromatogram and run log The result can be also viewed by using the BMP result file that is generated and stored on the USB memory stick For details refer to Section 6 6 3 BMP result file on page 208 Detailed instructions for the transfer of the result are presented in Section 6 6 Manage methods and files on page 195 Detailed instructions for the evaluation of a result are presented in Chapter 7 Operations from UNICORN start on page 210 and in the UNICORN start 1 0 User Manual Clean the system 192 After a run is completed perform the following e Remove the column from the flow path and re connect the flow path For detailed instructions see Section 8 3 1 Disconnect the column on page 223 e Rinse the flow path with cleaning solution and or DM water using either System cleaning or the Pump wash methods as required For detailed instructions see Section 8 3 Cleaning the system flow path on page 222 e f required remove the tubes from the Fraction collector If there is any spillage clean the Bowl assembly with DM water e Clean all spills on the instrument and on the bench using a damp cloth AKTA start Operating Instructions 29 0270 57 AG 6 Operations from the Instrument Display 6 5 Procedures after a run e Empty the waste container Clean and store the column After a run is completed perform the following e Disconnect the column from the flow path e Clean the column off line accor
186. ting Instructions 29 0270 57 AG 6 Operations from the Instrument Display 6 3 Perform a manual run 6 3 2 Monitor and control the run End the run Follow the instruction below to end an ongoing run Step Action 1 In the Run view screen tap End to terminate the run Run view UV absorbance 1 0 mAU Conductivity 25 0 mS cm Flow rate 3 0 ml min Graph Pressure 0 30 MPa Tube no 6 Edit run End Result A Message screen that requires to confirm the action opens Message A Are you sure you want to end the run Yes No Tap Yes to confirm that you want to terminate the run or tap No to cancel the action and return to the Run view screen Note When ending a run before it is completed the partial result is stored on the USB memory Stick The USB memory stick stores the result files which can be viewed using UNICORN start Also a BMP file is generated which can be viewed from any PC For details refer to Section 6 6 3 BMP result file on page 208 Do not remove the USB memory stick until the system generates the report BMP file Run ended Saal UV absorbance 1 0 mAU Conductivity 25 0 mS cm ee Flow rate 0 0 mi min Graph Pressure 0 30 MPa Tube no 6 Exit Tap Exit to close the Run ended screen AKTA start Operating Instructions 29 0270 57 AG 169 6 Operations from the Instrument Display 6 4 Perform a method run 6 4 Perform a method run Introduct
187. tor The following maintenance should be performed by the user of KTA start when required See section Section 8 6 2 Cleaning the instrument externally on page 232 Section 8 6 3 Cleaning the Fraction collector on page 2335 Perform System cleaning Section 8 3 2 System cleaning on page 224 Clean the UV flow cell Section 8 4 Cleaning the UV flow cell on page 227 Clean the Conductivity cell Section 8 5 Cleaning the Conductivity flow cell on page 229 Calibrate the touch screen Section 5 3 2 Instrument Display calibration on page 90 Calibrate the UV flow cell Section 5 3 5 UV Monitor calibration on page 96 Calibrate the Conductivity cell Section 5 3 6 Conductivity Monitor calibration on page 100 Pressure sensor zero offset Replace the inlet filters Section 5 3 3 Pressure sensor zero offset on page 92 Section 8 8 1 Replace the inlet filters on page 239 Replace the tubing and connec tors Section 8 8 2 Replace the tubing and connectors on page 240 AKTA start Operating Instructions 29 0270 57 AG 8 Maintenance 8 2 Cleaning before planned maintenance service 8 2 Cleaning before planned maintenance service Cleaning before planned maintenance service To ensure the protection and safety of service personnel all equipment and work areas must be clean and free of any hazardous contaminants before a Service Engineer starts maintenance work Please complete
188. tructions 29 0270 57 AG 209 7 Operations from UNICORN start 7 Operations from UNICORN start About this chapter This chapter gives a brief insight into the four modules of UNICORN start System Control Method Editor Evaluation and Administration In this chapter This chapter contains the following sections Section See page 7 1 System Control 211 7 2 Method Editor 213 7 3 Evaluation 215 7 4 Administration 216 Introduction UNICORN start brings the following capabilities e Simple and flexible method creation e Easy system control using process picture and real time monitoring of manual and method runs e Ability to evaluate and compare results e Create and print PDF reports e Ability to manage store archive retrieve backup restore results generated from AKTA start For detailed instructions on operations from UNICORN start refer to the UNICORN start 1 0 User Manual 210 AKTA start Operating Instructions 29 0270 57 AG 7 Operations from UNICORN start 7 1 System Control 7 1 System Control Introduction The System Control module is used to start view and control a run Illustration of the System Control user interface cr l E System Control 2 8 xs File View Manual System Tools Help an Manual Run Method Run Connect P Disconnect fh Eo C AKTA start Chromatogram x Conductivity Gradient Concentration System Flow Pressur
189. ual run of 50 B for 10 min can also be performed to evaluate the gradient fluctuations wavy gradients Illustration of a typical operation The illustration below shows the result of a System performance method where fluctu ations were observed in the gradient arrow 180 4 UV 4 Conductivity 4 Gradient Concentration System Flow Pressure Fraction cores Run Log AKTA start Operating Instructions 29 0270 57 AG 117 5 Prepare the system for a run 5 4 System performance 5 4 4 Switch valve timing The Switch valve timing was changed to 5 sec the test was repeated and the result showed an acceptable gradient performance arrow 4 UV 4 Conductivity 4 Gradient Concentration O System Flow O Pressure Fraction O Run Log 180 120 o8 88s Note Alternatively a manual run of 50 B for 10 min can also be performed to evaluate the gradient fluctuations wavy gradients Set Switch valve timing Follow the instructions below to change the Switch valve timing The Switch valve timing can be changed from the Instrument Display Step Action 1 From the AKTA start home screen tap Settings and service Result The Settings and service screen 1 opens Settings and service Fraction collector Pressure sensor 1 3 ET Next 118 KTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 4
190. uctions 29 0270 57 AG 6 Operations from the Instrument Display 6 3 Perform a manual run 6 3 2 Monitor and control the run 6 3 2 Monitor and control the run Overview From the Run view screen the user can monitor and control the ongoing run Run view UV absorbance 1 0 mAU Conductivity 25 0 mS cm Flow rate 3 0 ml min Graph Pressure 0 30 MPa Tube no 6 4 Edit run Pause f End The following options are available Description Graph Displays the run time UV absorbance curve Edit run Allows the user to edit the run parameters of the ongoing run Pause Temporarily pauses the run by stopping the Pump hence no flow of liquid in the flow path End Terminates the current run View the chromatogram Follow the instruction below to view the chromatogram of the ongoing run Step Action 1 Run view UV absorbance 1 0 mAU Conductivity 25 0 mS cm Flow rate 3 0 ml min Graph Pressure 0 30 MPa Tube no 6 Edit run Pause End In the Run view screen tap the graph icon to view the chromatogram KTA start Operating Instructions 29 0270 57 AG 165 6 Operations from the Instrument Display 6 3 Perform a manual run 6 3 2 Monitor and control the run Step Action 3 5 z E 2 z 5 The graph displays the UV curve The Y axis displays the UV absorbance MAU and X axis the time min Tap th
191. uctivity to Outlet valve PEEK tubing Length 19 cm i d 0 50 mm 10 32 UNF connections Outlet valve to Frac30 PEEK tubing Length 50 cm i d 0 75 mm 10 32 UNF connections Waste tubing ETFE tubing Length 60 cm i d 1 0 mm Fingertight connector 1 16 Sample tubing ETFE tubing Length 25 cm i d 1 0 mm Fingertight connector 1 16 Equipment noise level Parameter Value Environmental ranges Parameter Operation site Indoor use Altitude Maximum 2000 m Storage and transport temperature range 25 C to 60 C Humidity 20 to 80 non condensing Pollution degree 2 260 AKTA start Operating Instructions 29 0270 57 AG 10 Reference information 10 1 Specifications Operating range Parameter Operating temperature range 4 C to 35 C Relative humidity 20 to 80 non condensing Pump Parameter Pump type Peristaltic Pump Single channel four roller pump head with low pulsation Flow rate 0 5 to 5 ml min operating range 10 ml min Wash flow Flow rate specifications e Accuracy Flow rate lt 1 ml min 15 Flow rate gt 1 ml min 10 e Precision Flow rate lt 1 ml min 15 Flow rate gt 1 ml min 10 Condition 0 8 to 2 cP and fresh Pump tubing Pressure range O to 0 5 MPa 5 Bar Viscosity range 0 6 to 5 cP Mixer Parameter Mixing principle Static mixer Mixer volume 0 4 ml KTA start Operating Instructions 2
192. uick start e ACstep1mlHiTrap e AC step 5 ml HiTrap e DS5mlHiTrap e DS 53 ml HiPrep e IEX step 1 ml HiTrap e IEX step 5 ml HiTrap e IEX gradient 1 ml HiTrap e IEX gradient 5 ml HiTrap e GF 16 60 HiPrep Templates e Affinity AC e Desalting buffer exchange DS e lon exchange IEX e Gel filtration GF User defined Methods created by the user based on the predefined templates AKTA start Operating Instructions 29 0270 57 AG 171 6 Operations from the Instrument Display 6 4 Perform a method run 6 4 1 Select a method type Method type Prepare system Pump wash A Pump Wash B Washout fractionation tubing Column preparation System cleaning System performance method Select a method Follow the instruction below to select a method Step Action il KTA start Method run Manual run Create method Settings and service In the AKTA start home screen tap Method run to access the method types available with the instrument Method run 2 Quick start Templates User defined Prepare system Select one of the following methods e Quick start e Templates e User defined e Prepare system 172 KTA start Operating Instructions 29 0270 57 AG 6 Operations from the Instrument Display 6 4 Perform a method run 6 4 2 Quick start 6 4 2 Quick start Introduction Quick start contains ready to run methods to purify most common proteins based on Affinity lon exchan
193. ver to the left Load Sample position or the the right Inject to column position LOAD INJECT LOAD INJECT Function Default route for the system flow path Directs the liquid manually injected through port 3 to the sample loop Note A sample loop or a Superloop is connected to the ports 2 and 5 of the Injection valve Directs the liquid from the sample loop through port 4 to the waste container Note The path indicated by light orange arrows in the illustration above is used during the manual filling of the loop Sample or Superloop through port 3 136 AKTA start Operating Instructions 29 0270 57 AG 5 Prepare the system for a run 5 7 Sample application Valve position Inject to column Port connection Function Diverts the system flow path to the sample loop Directs the liquid from the sample loop to the column so that the sample loaded into the loop is transferred to the column Connect a sample loop Follow the instructions below to connect a sample loop to the Injection valve Step Action 1 Connect the sample loop between ports 2 and 5 of the Injection valve 2 Make sure that waste tubing is connected to port 4 of the Injection valve Connect a Superloop Follow the instructions below to connect a Superloop to the Injection valve Step Action 1 Attach an appropriate column holder to the column holder rail on the right side edge of the instrument Z Make sure
194. w to operate the instrument perform a run and the procedures after a run In this chapter This chapter contains the following sections Section See page 6 1 Introduction 156 6 2 Fractionation 159 6 3 Perform a manual run 162 6 4 Perform a method run 170 6 5 Procedures after a run 192 6 6 Manage methods and files 195 155 KTA start Operating Instructions 29 0270 57 AG 6 Operations from the Instrument Display 6 1 Introduction 6 1 Introduction Workflow A typical workflow for AKTA start is presented below All the operations in the workflow can be controlled from the instrument display A chromatographic run can be performed by either using a Quick start method Template or by operating the system manually The options for starting a run from the instrument display are e Method run e Manual run Detailed instructions are presented in Section 6 3 Perform a manual run on page 162 and Section 6 4 Perform a method run on page 170 KTA start home Method run Create method Settings and service Method run options Manual run Create method options Set run parameters F Create Edit USB import e Quick start L Delete Settings and service options for the modules e Templates e User defined e Prepare system e Fraction collector e Pressure sensor e Pump e System Save new method e Buffer valve e Sample valve e Wash valve e Outlet valve User defined e Main board i UV e Disp
195. y during the predefined cleaning procedure Pump wash In a manual run the valve can be set to the intended position by configuring the run parameters Injection valve A 6 port manually operated valve that is used to transfer the sample loaded in the sample loop on to the column A sample loop is connected to the appropriate ports of the valve The valve is switched manually to positions e Load sample default to allow the loading of the sample into the sample loop e Inject to column to transfer the sample from the loop on to the column during a chromatography run UV The UV Monitor continuously measures the ab sorbance of the liquid in the UV flow cell at a set wavelength of 280 nm The UV flow cell has a path length of 2 mm AKTA start Operating Instructions 29 0270 57 AG 39 3 System description 3 2 Instrument Function Description Conductivity The Conductivity Monitor continuously reads the conductivity of the liquid in the Conductivity flow cell The conductivity is automatically calculated by multiplying the measured conductance by the cell constant of the flow cell The cell constant is factory calibrated The Conductivity flow cell is provided with a temperature sensor that measures the tempera ture of the liquid in the Conductivity flow cell Note The buffers used should be within the conductivity range of the instrument 0 to 300 mS cm Outlet valve A 3 port valve tha
196. y requirements of EN IEC 61010 1 or relevant harmonized standards Within EU connected equipment must be CE marked 10 KTA start Operating Instructions 29 0270 57 AG 1 Introduction 1 3 Regulatory information Environmental conformity This product conforms to the following environmental requirements Requirement 2011 65 EU Restriction of Hazardous Substances ROHS Directive 2012 19 EU Waste Electrical and Electronic Equipment WEEE Directive ACPEIP Administration on the Control of Pollution Caused by Electronic Information Products China Restriction of Hazardous Sub stances ROHS Regulation EC Registration Evaluation Authorization and restriction of No 1907 2006 CHemicals REACH AKTA start Operating Instructions 29 0270 57 AG 11 1 Introduction 1 4 Associated documentation 14 Associated documentation Introduction This section describes the user documentation that is delivered with the instrument and how to find related literature that can be downloaded or ordered from GE User documentation on the CD The user documentation listed in the table below is available on the AKTA start User Documentation CD A display help is available from the Instrument Display Documentation KTA start Operating Instructions Main contents Instructions needed to install operate and maintain KTA start in a safe way KTA start Instrument Display Help Dialog descriptions of the functionality
197. ying to remove the tray from AKTA start e NaOH is corrosive and therefore dangerous to health When using hazardous chemicals avoid spillage and wear protective glasses and other suitable Personal Protective Equipment PPE CAUTION To avoid damage to person when performing maintenance on AKTA start follow the instructions below e Hazardous UV light Always switch off power to the instrument before replacing the UV flow cell e f hazardous chemicals are used for system or column cleaning wash the system or columns with a neutral solution in the last phase or step before maintenance 22 AKTA start Operating Instructions 29 0270 57 AG 2 Safety instructions 2 1 Safety precautions NOTICE Cleaning Keep the instrument dry and clean Wipe regularly with asoft damp tissue and if necessary a mild cleaning agent Let the instrument dry completely before use AKTA start Operating Instructions 29 0270 57 AG 23 2 Safety instructions 2 2 Labels 2 2 Labels Introduction This section describes the safety labels and labels concerning hazardous substances that are attached to AKTA start The instrument serial number is also visible from the instrument product label which is illustrated here Labels on AKTA start The illustrations below show the labels that are attached to AKTA start Description Placement 2902209 AKTA start Instrument label including S a Aa Fre Aane 50 60 He safet

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